LOCUS Exported 8182 bp ds-DNA circular SYN 22-APR-2014

DEFINITION .

ACCESSION .

VERSION .

KEYWORDS ZIP-SFFV-NTC1

SOURCE synthetic DNA construct

ORGANISM synthetic DNA construct

REFERENCE 1 (bases 1 to 8182)

AUTHORS Trial User

TITLE Direct Submission

JOURNAL Exported Thursday, May 01, 2014 from SnapGene 2.3.4

FEATURES Location/Qualifiers

source 1..8182

/organism="synthetic DNA construct"

/mol_type="other DNA"

misc_feature 23..667

/note="CMV-LTR"

misc_feature 720..856

/note="psi"

misc_feature 1348..1552

/note="rre"

misc_feature 2062..2212

/note="cts"

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 2634..3329

/codon_start=1

/label="ZsGreen"

/note="ZsGreen"

/translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN

LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV

CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG

DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH

AIASGSALP"

misc_feature 3341..3917

/note="IRES"

CDS 3918..4520

/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 4538..4632

/note="5'-miR30"

misc_feature 4628..4647

/note="Illumina actual seq primer"

misc_feature 4633..4748

/note="Internal Mir30 context with NTC1 shRNA"

misc_feature 4688..4706

/note="Loop"

misc_feature 4746..4856

/note="3'-miR30"

misc_feature 4887..5478

/note="WPRE"

misc_feature 5683..5918

/note="3'LTR"

promoter 6012..6341

/note="SV40"

/note="SV40 enhancer and early promoter"

rep_origin 6192..6327

/note="ori"

/note="SV40 origin of replication"

rep_origin complement(6430..7015)

/direction=LEFT

/note="ori"

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

replication"

CDS complement(7186..8145)

/gene="bla"

/product="beta-lactamase"

/note="AmpR"

/note="confers resistance to ampicillin, carbenicillin, and

related antibiotics"

ORIGIN

1 gacggatcgg gagatcctcg cgcgttgaca ttgattattg actagttatt aatagtaatc

61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt

121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta

181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg

241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga

301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt

361 tcctacttgg cagtacatct acgtattagt catcgctatt accagctagc gaggcgtggc

421 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc

481 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg

541 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg

601 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat

661 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct

721 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg

781 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc

841 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga

901 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca

961 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa

1021 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc

1081 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag

1141 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac

1201 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa

1261 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa

1321 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta

1381 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc

1441 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag

1501 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc

1561 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt

1621 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg

1681 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc

1741 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt

1801 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag

1861 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc

1921 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc

1981 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga

2041 acggatctcg acggtatcgc ctttaaaaga aaagggggga ttggggggta cagtgcaggg

2101 gaaagaatag tagacataat agcaacagac atacaaacta aagaactaca aaaacaaatt

2161 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttatcgataa

2221 gcttagctag ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata

2281 gagaagttca gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat

2341 ctgcggtgag cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg

2401 gccccggccc gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc

2461 ttaagaccca tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat

2521 ttgaattaac caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc

2581 tataaaagag ctcacaaccc ctcactcggc gcgccagtcc tccgaaccgg tccatggccc

2641 agtccaagca cggcctgacc aaggagatga ccatgaagta ccgcatggag ggctgcgtgg

2701 acggccacaa gttcgtgatc accggcgagg gcatcggcta ccccttcaag ggcaagcagg

2761 ccatcaacct gtgcgtggtg gagggcggcc ccttgccctt cgccgaggac atcttgtccg

2821 ccgccttcat gtacggcaac cgcgtgttca ccgagtaccc ccaggacatc gtcgactact

2881 tcaagaactc ctgccccgcc ggctacacct gggaccgctc cttcctgttc gaggacggcg

2941 ccgtgtgcat ctgcaacgcc gacatcaccg tgagcgtgga ggagaactgc atgtaccacg

3001 agtccaagtt ctacggcgtg aacttccccg ccgacggccc cgtgatgaag aagatgaccg

3061 acaactggga gccctcctgc gagaagatca tccccgtgcc caagcagggc atcttgaagg

3121 gcgacgtgag catgtacctg ctgctgaagg acggtggccg cttgcgctgc cagttcgaca

3181 ccgtgtacaa ggccaagtcc gtgccccgca agatgcccga ctggcacttc atccagcaca

3241 agctgacccg cgaggaccgc agcgacgcca agaaccagaa gtggcacctg accgagcacg

3301 ccatcgcctc cggctccgcc ttgccctgag aattcaattc cgcccctctc cctccccccc

3361 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt

3421 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt

3481 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa

3541 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac

3601 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg

3661 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt

3721 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca

3781 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt

3841 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg

3901 aaaaacacga taataccatg gccaccgagt acaagcccac ggtgcgcctc gccacccgcg

3961 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc

4021 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc

4081 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg

4141 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggctcgc

4201 gcatggccaa gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg

4261 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc

4321 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg

4381 ctggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc

4441 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga

4501 cccgcaagcc cggtgcctga gcggccgcaa tggatcttgt ttgaatgagg cttcagtact

4561 ttacagaatc gttgcctgca catcttggaa acacttgctg ggattacttc ttcaggttaa

4621 cccaacagaa ggctcgagaa ggtatattgc tgttgacagt gagcgcccgg ctgaagagcc

4681 tgatcaatag tgaagccaca gatgtattga tcaggctctt cagccggttg cctactgcct

4741 cggaattcaa ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc

4801 tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt

4861 aattaaacgc gtggcgcgcc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat

4921 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc

4981 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct

5041 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca

5101 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt

5161 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg

5221 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga

5281 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt

5341 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc

5401 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt

5461 gggccgcctc cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat

5521 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga

5581 ggaggaggtg ggtttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg

5641 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact

5701 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct

5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc

5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc

5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta

5941 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgtt

6001 taaactggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa

6061 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc

6121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc

6181 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct

6241 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga

6301 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta

6361 tatccattac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt

6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag

6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc

6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc

6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt

6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt

6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc

6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa

6841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa

6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg

6961 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga

7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat

7081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag

7141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat

7201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc

7261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat

7321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag

7381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg

7441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc

7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca

7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg

7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc

7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta

7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc

7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg

7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc

7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc

7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat

8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag

8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc

8161 ccgaaaagtg ccacctgacg tc

//