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
//