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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