LOCUS       sequence-10068-e        5803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5803)
  AUTHORS   Kota J, Chivukula RR, O'Donnell KA, Wentzel EA, Montgomery CL, Hwang
            HW, Chang TC, Vivekanandan P, Torbenson M, Clark KR, Mendell JR, 
            Mendell JT
  TITLE     Therapeutic microRNA delivery suppresses tumorigenesis in a murine 
            liver cancer model.
  JOURNAL   Cell. 2009 Jun 12. 137(6):1005-17.
  PUBMED    19524505
REFERENCE   2  (bases 1 to 5803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009 
            Jun 12. 137(6):1005-17."
COMMENT     Sequence Label: scAAV GFP
FEATURES             Location/Qualifiers
     source          1..5803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        155..366
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          755..958
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     915..935
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    971..1051
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1053..1062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1059..1061,1062..1064,1065..1778)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1059..1061,1062..1064,1065..1775)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1104..1125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1365..1384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1712..1733
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1791..1925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1815..1834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1869..1888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2587..3100
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2674..2693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2884..2905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3581..3600)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3700..3722
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3760..3778)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3846..3950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3951..4019,4020..4811)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4169..4188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4982..5570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5471..5490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggggttatcg
      121 gcgcgccact agtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
      181 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      241 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      301 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      361 acacaggtaa gtgccgtgtg tggttcccgc gggcggcgac ggggcccgtg cgtcccagcg
      421 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
      481 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
      541 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
      601 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
      661 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
      721 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
      781 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
      841 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
      901 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
      961 gtcgtgaaaa gctagcgcta ccggactcag atctcgagct caagcttcga attctgcagt
     1021 cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1081 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1141 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1201 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1261 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1321 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1381 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1441 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1501 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1561 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1621 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1681 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1741 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcggg gatccagaca
     1801 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1861 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1921 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1981 tttttttcta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     2041 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2101 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2161 agcgcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2221 gcagcctgaa tggcgaatgg aattccagac gattgagcgt caaaatgtag gtatttccat
     2281 gagcgttttt cctgttgcaa tggctggcgg taatattgtt ctggatatta ccagcaaggc
     2341 cgatagtttg agttcttcta ctcaggcaag tgatgttatt actaatcaaa gaagtattgc
     2401 gacaacggtt aatttgcgtg atggacagac tcttttactc ggtggcctca ctgattataa
     2461 aaacacttct caggattctg gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct
     2521 gtttagctcc cgctctgatt ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac
     2581 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2641 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2701 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2761 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2821 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2881 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2941 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3001 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaatattt gcttatacaa
     3061 tcttcctgtt tttggggctt ttctgattat caaccggggt acatatgatt gacatgctag
     3121 ttttacgatt accgttcatc gattctcttg tttgctccag actctcaggc aatgacctga
     3181 tagcctttgt agagacctct caaaaatagc taccctctcc ggcatgaatt tatcagctag
     3241 aacggttgaa tatcatattg atggtgattt gactgtctcc ggcctttctc acccgtttga
     3301 atctttacct acacattact caggcattgc atttaaaata tatgagggtt ctaaaaattt
     3361 ttatccttgc gttgaaataa aggcttctcc cgcaaaagta ttacagggtc ataatgtttt
     3421 tggtacaacc gatttagctt tatgctctga ggctttattg cttaattttg ctaattcttt
     3481 gccttgcctg tatgatttat tggatgttgg aattcctgat gcggtatttt ctccttacgc
     3541 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3601 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3661 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3721 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3781 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3961 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4021 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4081 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4141 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4201 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4261 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4321 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4381 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4441 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4501 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4561 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4621 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4681 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4741 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4801 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4861 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4921 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4981 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5041 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5101 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5161 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5221 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5281 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5341 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5401 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5461 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5521 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5581 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5641 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5701 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5761 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atg
//
