LOCUS       sequence-126708-        6059 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 6059)
  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 6059)
  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: sc AAV miR26a eGFP
FEATURES             Location/Qualifiers
     source          1..6059
                     /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          1011..1214
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     1171..1191
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    1227..1307
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1309..1318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1315..1317,1318..1320,1321..2034)
                     /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(1315..1317,1318..1320,1321..2031)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1360..1381)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1621..1640)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1968..1989
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2047..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2125..2144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2843..3356
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2930..2949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3140..3161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3837..3856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3956..3978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4016..4034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4102..4206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4207..4275,4276..5067)
                     /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(4425..4444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5238..5826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5727..5746
                     /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 cggccccctg gtgcaattca ttacctaatc atgacctgga cagactgtcc
      541 tgtcggagcc aaggacagaa agctcccata gaggctgtgg ctggattcaa gtaatccagg
      601 ataggctgtt tccatctgtg aggcctattc ttgattactt gtttctggag gcagctgatg
      661 gtccgccgcc ggaaacagag atggctcctg ggacatggtg tgtgcgcttc ttcctgagcc
      721 aggttgaggt tgggaccact gatggccggc ctgctctggt gcctggcctc gcgccgccgt
      781 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
      841 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
      901 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
      961 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     1021 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     1081 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     1141 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     1201 ttcttccatt tcaggtgtcg tgaaaagcta gcgctaccgg actcagatct cgagctcaag
     1261 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg
     1321 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1381 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1441 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1501 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1561 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1621 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1681 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1741 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1801 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1861 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1921 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1981 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     2041 cgcggggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2101 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2161 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2221 ggggaggtgt gggaggtttt tttctagagc atggctacgt agataagtag catggcgggt
     2281 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2341 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2401 cctcagtgag cgagcgagcg cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2461 ccttcccaac agttgcgcag cctgaatggc gaatggaatt ccagacgatt gagcgtcaaa
     2521 atgtaggtat ttccatgagc gtttttcctg ttgcaatggc tggcggtaat attgttctgg
     2581 atattaccag caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta
     2641 atcaaagaag tattgcgaca acggttaatt tgcgtgatgg acagactctt ttactcggtg
     2701 gcctcactga ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc
     2761 ctttaatcgg cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg
     2821 tgctcgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2881 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2941 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3001 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3061 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3121 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3181 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3241 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa
     3301 atatttgctt atacaatctt cctgtttttg gggcttttct gattatcaac cggggtacat
     3361 atgattgaca tgctagtttt acgattaccg ttcatcgatt ctcttgtttg ctccagactc
     3421 tcaggcaatg acctgatagc ctttgtagag acctctcaaa aatagctacc ctctccggca
     3481 tgaatttatc agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc
     3541 tttctcaccc gtttgaatct ttacctacac attactcagg cattgcattt aaaatatatg
     3601 agggttctaa aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac
     3661 agggtcataa tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta
     3721 attttgctaa ttctttgcct tgcctgtatg atttattgga tgttggaatt cctgatgcgg
     3781 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3841 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3901 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3961 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4021 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4441 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4741 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5941 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6001 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatg
//
