LOCUS Exported 8211 bp ds-DNA circular SYN 14-JUL-2014

DEFINITION .

ACCESSION .

VERSION .

KEYWORDS pZIP-SFFV-RFP_miR30

SOURCE synthetic DNA construct

ORGANISM recombinant plasmid

REFERENCE 1 (bases 1 to 8211)

AUTHORS Transomic

TITLE Direct Submission

JOURNAL Exported Monday, Jul 21, 2014 from SnapGene 2.4.3

FEATURES Location/Qualifiers

source 1..8211

/organism="recombinant plasmid"

/mol_type="other DNA"

misc_feature 23..667

/note="CMV-LTR"

misc_feature 715..840

/note="HIV-1 Psi"

/note="packaging signal of human immunodeficiency virus

type 1"

misc_feature 1337..1570

/note="RRE"

/note="The Rev response element (RRE) of HIV-1 allows for

Rev-dependent mRNA export from the nucleus to the

cytoplasm."

misc_feature 2062..2178

/note="cPPT/CTS"

/note="central polypurine tract and central termination

sequence of HIV-1 (lacking the first T)"

misc_feature 2213..2214

/note="MCS"

promoter 2225..2625

/gene="SFFV"

/label="SFFV"

/note="SFFV"

primer_bind 2225..2625

/note="seq.ZsGrev"

primer_bind 2488..2625

/note="seq.SFFVfw"

CDS 2656..3351

/codon_start=1

/product="red fluorescent protein from Entacmaea

quadricolor"

/note="TurboRFP"

/note="mammalian codon-optimized"

/translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV

VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA

TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK

LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC

DLPSKLGHR"

misc_feature 3363..3939

/note="IRES"

CDS 3940..4542

/codon_start=1

/gene="pac from Streptomyces alboniger"

/product="puromycin N-acetyltransferase"

/note="PuroR"

/note="confers resistance to puromycin"

/translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE

RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAKLSGSRL

AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET

SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"

misc_feature 4566..4661

/note="5' mir30"

misc_feature 4566..4594

/note="Seq Primer -1 97 miR"

misc_feature 4566..4573

/note="partial forward 5' illumina adapted primer"

misc_feature 4656..4693

/note="Illumina actual seq primer"

misc_feature 4683..4767

/note="miR30 shRNA"

gap 4694..4715

/estimated_length="22"

gap 4735..4756

/estimated_length="22"

misc_feature 4772..4884

/note="3' mir30"

misc_feature 4916..5504

/note="WPRE"

/note="woodchuck hepatitis virus posttranscriptional

regulatory element"

LTR 5712..5945

/note="3' LTR (Delta-U3)"

/note="self-inactivating 3' long terminal repeat (LTR)fromHIV-1"

promoter 6041..6370

/note="SV40"

/note="SV40 enhancer and early promoter"

rep_origin 6221..6356

/note="ori"

/note="SV40 origin of replication"

rep_origin complement(6459..7044)

/direction=LEFT

/note="ori"

/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

replication"

CDS complement(7215..8174)

/gene="bla"

/product="beta-lactamase"

/note="AmpR"

/note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"

promoter complement(8076..8180)

/gene="bla"

/note="AmpR promoter"

ORIGIN

1 gacggatcgggagatcctcgcgcgttgacattgattattgactagttattaatagtaatc

61 aattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggt

121 aaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgta

181 tgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacg

241 gtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattga

301 cgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactt

361 tcctacttggcagtacatctacgtattagtcatcgctattaccagctagcgaggcgtggc

421 ctgggcgggactggggagtggcgagccctcagatcctgcatataagcagctgctttttgc

481 ctgtactgggtctctctggttagaccagatctgagcctgggagctctctggctaactagg

541 gaacccactgcttaagcctcaataaagcttgccttgagtgcttcaagtagtgtgtgcccg

601 tctgttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaaaat

661 ctctagcagtggcgcccgaacagggacttgaaagcgaaagggaaaccagaggagctctct

721 cgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcggcgactggtg

781 agtacgccaaaaattttgactagcggaggctagaaggagagagatgggtgcgagagcgtc

841 agtattaagcgggggagaattagatcgcgatgggaaaaaattcggttaaggccaggggga

901 aagaaaaaatataaattaaaacatatagtatgggcaagcagggagctagaacgattcgca

961 gttaatcctggcctgttagaaacatcagaaggctgtagacaaatactgggacagctacaa

1021 ccatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccctc

1081 tattgtgtgcatcaaaggatagagataaaagacaccaaggaagctttagacaagatagag

1141 gaagagcaaaacaaaagtaagaccaccgcacagcaagcggccggccgctgatcttcagac

1201 ctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagtagtaa

1261 aaattgaaccattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaa

1321 aaagagcagtgggaataggagctttgttccttgggttcttgggagcagcaggaagcacta

1381 tgggcgcagcgtcaatgacgctgacggtacaggccagacaattattgtctggtatagtgc

1441 agcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaactcacag

1501 tctggggcatcaagcagctccaggcaagaatcctggctgtggaaagatacctaaaggatc

1561 aacagctcctggggatttggggttgctctggaaaactcatttgcaccactgctgtgcctt

1621 ggaatgctagttggagtaataaatctctggaacagatttggaatcacacgacctggatgg

1681 agtgggacagagaaattaacaattacacaagcttaatacactccttaattgaagaatcgc

1741 aaaaccagcaagaaaagaatgaacaagaattattggaattagataaatgggcaagtttgt

1801 ggaattggtttaacataacaaattggctgtggtatataaaattattcataatgatagtag

1861 gaggcttggtaggtttaagaatagtttttgctgtactttctatagtgaatagagttaggc

1921 agggatattcaccattatcgtttcagacccacctcccaaccccgaggggacccgacaggc

1981 ccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattagtga

2041 acggatctcgacggtatcgcctttaaaagaaaaggggggattggggggtacagtgcaggg

2101 gaaagaatagtagacataatagcaacagacatacaaactaaagaactacaaaaacaaatt

2161 acaaaaattcaaaattttcgggtttattacagggacagcagagatccagtttatcgattt

2221 ggcaagctagctgcagtaacgccattttgcaaggcatggaaaaataccaaaccaagaata

2281 gagaagttcagatcaagggcgggtacatgaaaatagctaacgttgggccaaacaggatat

2341 ctgcggtgagcagtttcggccccggcccggggccaagaacagatggtcaccgcagtttcg

2401 gccccggcccgaggccaagaacagatggtccccagatatggcccaaccctcagcagtttc

2461 ttaagacccatcagatgtttccaggctcccccaaggacctgaaatgaccctgcgccttat

2521 ttgaattaaccaatcagcctgcttctcgcttctgttcgcgcgcttctgcttcccgagctc

2581 tataaaagagctcacaacccctcactcggcgcgccagtcctccgacagactgagtcgccc

2641 gggaccggtgccaccatgagcgagctgatcaaggagaacatgcacatgaagctgtacatg

2701 gagggcaccgtgaacaaccaccacttcaagtgcacatccgagggcgaaggcaagccctac

2761 gagggcacccagaccatgaagatcaaggtggtcgagggcggccctctccccttcgccttc

2821 gacatcctggctaccagcttcatgtacggcagcaaagccttcatcaaccacacccagggc

2881 atccccgacttctttaagcagtccttccctgagggcttcacatgggagagaatcaccaca

2941 tacgaagacgggggcgtgctgaccgctacccaggacaccagcttccagaacggctgcatc

3001 atctacaacgtcaagatcaacggggtgaacttcccatccaacggccctgtgatgcagaag

3061 aaaacacgcggctgggaggccaacaccgagatgctgtaccccgctgacggcggcctgaga

3121 ggccacagccagatggccctgaagctcgtgggcgggggctacctgcactgctccttcaag

3181 accacatacagatccaagaaacccgctaagaacctcaagatgcccggcttccacttcgtg

3241 gaccacagactggaaagaatcaaggaggccgacaaagagacctacgtcgagcagcacgag

3301 atggctgtggccaagtactgcgacctccctagcaaactggggcacagataagaattcaat

3361 tccgcccctctccctcccccccccctaacgttactggccgaagccgcttggaataaggcc

3421 ggtgtgcgtttgtctatatgttattttccaccatattgccgtcttttggcaatgtgaggg

3481 cccggaaacctggccctgtcttcttgacgagcattcctaggggtctttcccctctcgcca

3541 aaggaatgcaaggtctgttgaatgtcgtgaaggaagcagttcctctggaagcttcttgaa

3601 gacaaacaacgtctgtagcgaccctttgcaggcagcggaaccccccacctggcgacaggt

3661 gcctctgcggccaaaagccacgtgtataagatacacctgcaaaggcggcacaaccccagt

3721 gccacgttgtgagttggatagttgtggaaagagtcaaatggctctcctcaagcgtattca

3781 acaaggggctgaaggatgcccagaaggtaccccattgtatgggatctgatctggggcctc

3841 ggtgcacatgctttacatgtgtttagtcgaggttaaaaaaacgtctaggccccccgaacc

3901 acggggacgtggttttcctttgaaaaacacgataataccatggccaccgagtacaagccc

3961 acggtgcgcctcgccacccgcgacgacgtcccccgggccgtacgcaccctcgccgccgcg

4021 ttcgccgactaccccgccacgcgccacaccgtcgacccggaccgccacatcgagcgggtc

4081 accgagctgcaagaactcttcctcacgcgcgtcgggctcgacatcggcaaggtgtgggtc

4141 gcggacgacggcgccgcggtggcggtctggaccacgccggagagcgtcgaagcgggggcg

4201 gtgttcgccgagatcggctcgcgcatggccaagttgagcggttcccggctggccgcgcag

4261 caacagatggaaggcctcctggcgccgcaccggcccaaggagcccgcgtggttcctggcc

4321 accgtcggcgtctcgcccgaccaccagggcaagggtctgggcagcgccgtcgtgctcccc

4381 ggagtggaggcggccgagcgcgctggggtgcccgccttcctggagacctccgcgccccgc

4441 aacctccccttctacgagcggctcggcttcaccgtcaccgccgacgtcgaggtgcccgaa

4501 ggaccgcgcacctggtgcatgacccgcaagcccggtgcctgagcggccgcaatggatctc

4561 tcgactgtttgaatgaggcttcagtactttacagaatcgttgcctgcacatcttggaaac

4621 acttgctgggattacttcttcaggttaacccaacagaaggctcgagaaggtatattgctg

4681 ttgacagtgagcgnnnnnnnnnnnnnnnnnnnnnntagtgaagccacagatgtannnnnn

4741 nnnnnnnnnnnnnnnntgcctactgcctcggaattcaaggggctactttaggagcaatta

4801 tcttgtttactaaaactgaataccttgctatctctttgatacatttttacaaagctgaat

4861 taaaatggtataaattaaatcacttttttcctaggacgcgtggcgcgcctggaacaatca

4921 acctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgctccttt

4981 tacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttcccgtatggc

5041 tttcattttctcctccttgtataaatcctggttgctgtctctttatgaggagttgtggcc

5101 cgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttg

5161 gggcattgccaccacctgtcagctcctttccgggactttcgctttccccctccctattgc

5221 cacggcggaactcatcgccgcctgccttgcccgctgctggacaggggctcggctgttggg

5281 cactgacaattccgtggtgttgtcggggaagctgacgtcctttccatggctgctcgcctg

5341 tgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcggccctcaatcc

5401 agcggaccttccttcccgcggcctgctgccggctctgcggcctcttccgcgtcttcgcct

5461 tcgccctcagacgagtcggatctccctttgggccgcctccccgcctggaattaattctgc

5521 agtcgagacctagaaaaacatggagcaatcacaagtagcaatacagcagctaccaatgct

5581 gattgtgcctggctagaagcacaagaggaggaggaggtgggtttttccagtcacacctca

5641 ggtacctttaagaccaatgacttacaaggcagctgtagatcttagccactttttaaaaga

5701 aaagaggggactggaagggctaattcactcccaacgaagacaagatctgctttttgcttg

5761 tactgggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaa

5821 cccactgcttaagcctcaataaagcttgccttgagtgcttcaagtagtgtgtgcccgtct

5881 gttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaaaatctc

5941 tagcagtagtagttcatgtcatcttattattcagtatttataacttgcaaagaaatgaat

6001 atcagagagtgagaggccttgacattgtttaaactggaatgtgtgtcagttagggtgtgg

6061 aaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagc

6121 aaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatct

6181 caattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcc

6241 cagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccga

6301 ggccgcctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctagg

6361 cttttgcaaaaagctcccgggagcttgtatatccattacatgtgagcaaaaggccagcaa

6421 aaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccct

6481 gacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataa

6541 agataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccg

6601 cttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctca

6661 cgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaa

6721 ccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccg

6781 gtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgagg

6841 tatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaga

6901 acagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagc

6961 tcttgatccggcaaacaaaccaccgctggtagcggtttttttgtttgcaagcagcagatt

7021 acgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgct

7081 cagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttc

7141 acctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaa

7201 acttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtcta

7261 tttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggc

7321 ttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagat

7381 ttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaacttta

7441 tccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagtt

7501 aatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgttt

7561 ggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatg

7621 ttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggcc

7681 gcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatcc

7741 gtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatg

7801 cggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcaga

7861 actttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatctta

7921 ccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatct

7981 tttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaag

8041 ggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattga

8101 agcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaat

8161 aaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgt c

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