LOCUS       sequence-415446-        5331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre by the specific cTNT promoter and incorporation of the
            miR122 target sequences (miR122TS) into the 3' untranslated region 
            (3' UTR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5331)
  AUTHORS   Yang L, Liu Z, Chen G, Chen Z, Guo C, Ji X, Cui Q, Sun Y, Hu X, 
            Zheng Y, Li Y, Gao F, Chen L, Zhou P, Pu WT, Guo Y
  TITLE     MicroRNA-122-Mediated Liver Detargeting Enhances the Tissue 
            Specificity of Cardiac Genome Editing.
  JOURNAL   Circulation. 2024 May 28;149(22):1778-1781. doi: 
            10.1161/CIRCULATIONAHA.123.065438. Epub 2024 May 28.
  PUBMED    38805581
REFERENCE   2  (bases 1 to 5331)
  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"; doi: 
            "10.1161/CIRCULATIONAHA.123.065438"; journalName: "Circulation"; 
            date: "2024-05-28- 28"; volume: "149"; issue: "22"; pages: 
            "1778-1781"
COMMENT     Sequence Label: pAAV-cTNT-Cre-miR122 TS
FEATURES             Location/Qualifiers
     source          1..5331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(300..319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     419..441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(479..497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        565..669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(670..738,739..1530)
                     /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(888..907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1701..2289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2190..2209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2351..2480
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          3041..3173
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3156..3175
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3218..3236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3248..3268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4428..4904
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4627..4646)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4944..5084
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4944..5073
                     /label=AAV2 ITR
     rep_origin      join(5159..5331,1..283)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5246..5265)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
       61 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      121 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
      181 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
      241 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
      301 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
      481 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2341 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     2401 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     2461 ctccatcact aggggttcct gcggccgcac gcgccggtac cgggataaaa gcagtctggg
     2521 ctttcacatg acagcatctg gggctgcggc agagggtcgg gtccgaagcg ctgccttatc
     2581 agcgtcccca gccctgggag gtgacagctg gctggcttgt gtcagcccct cgggcactca
     2641 cgtatctccg tccgacgggt ttaaaatagc aaaactctga ggccacacaa tagcttgggc
     2701 ttatatgggc tcctgtgggg gaagggggag cacggagggg gccggggccg ctgctgccaa
     2761 aatagcagct cacaagtgtt gcattcctct ctgggcgccg ggcacattcc tgctggctct
     2821 gcccgccccg gggtgggcgc cggggggacc ttaaagcctc tgccccccaa ggagcccttc
     2881 ccagacagcc gccggcaccc accgctccgt gggacgatcc ccgaagctct agagctttat
     2941 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc
     3001 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
     3061 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
     3121 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
     3181 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
     3241 catggtgccc aagaagaaga ggaaagtctc caacctgctg actgtgcacc aaaacctgcc
     3301 tgccctccct gtggatgcca cctctgatga agtcaggaag aacctgatgg acatgttcag
     3361 ggacaggcag gccttctctg aacacacctg gaagatgctc ctgtctgtgt gcagatcctg
     3421 ggctgcctgg tgcaagctga acaacaggaa atggttccct gctgaacctg aggatgtgag
     3481 ggactacctc ctgtacctgc aagccagagg cctggctgtg aagaccatcc aacagcacct
     3541 gggccagctc aacatgctgc acaggagatc tggcctgcct cgcccttctg actccaatgc
     3601 tgtgtccctg gtgatgagga gaatcagaaa ggagaatgtg gatgctgggg agagagccaa
     3661 gcaggccctg gcctttgaac gcactgactt tgaccaagtc agatccctga tggagaactc
     3721 tgacagatgc caggacatca ggaacctggc cttcctgggc attgcctaca acaccctgct
     3781 gcgcattgcc gaaattgcca gaatcagagt gaaggacatc tcccgcaccg atggtgggag
     3841 aatgctgatc cacattggca ggaccaagac cctggtgtcc acagctggtg tggagaaggc
     3901 cctgtccctg ggggttacca agctggtgga gagatggatc tctgtgtctg gtgtggctga
     3961 tgaccccaac aactacctgt tctgccgggt cagaaagaat ggtgtggctg ccccttctgc
     4021 cacctcccaa ctgtccaccc gggccctgga agggatcttt gaggccaccc accgcctgat
     4081 ctatggtgcc aaggatgact ctgggcagag atacctggcc tggtctggcc actctgccag
     4141 agtgggtgct gccagggaca tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc
     4201 tggtggctgg accaatgtga acattgtgat gaactacatc agaaacctgg actctgagac
     4261 tggggccatg gtgaggctgc tcgaggatgg ggactgaaag cttcggtcaa acaccattgt
     4321 cacactccat actatcaaac accattgtca cactccagat caacaaacac cattgtcaca
     4381 ctccacgccg aagcttatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     4441 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     4501 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     4561 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     4621 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     4681 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     4741 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     4801 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     4861 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4921 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4981 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     5041 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     5101 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     5161 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5221 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5281 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt c
//
