LOCUS       sequence-43859-e        5915 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 5915)
  AUTHORS   Xie J, Ameres SL, Friedline R, Hung JH, Zhang Y, Xie Q, Zhong L, Su 
            Q, He R, Li M, Li H, Mu X, Zhang H, Broderick JA, Kim JK, Weng Z, 
            Flotte TR, Zamore PD, Gao G
  TITLE     Long-term, efficient inhibition of microRNA function in mice using 
            rAAV vectors.
  JOURNAL   Nat Methods. 2012 Mar 4;9(4):403-9. doi: 10.1038/nmeth.1903.
  PUBMED    22388288
REFERENCE   2  (bases 1 to 5915)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-03-4- 4"; volume: "9"; 
            issue: "4"; pages: "403-9"
COMMENT     Sequence Label: pAAV TBG FFLuc
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        481..890
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     CDS             1003..2655
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1069..1087)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2753..2772)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2818..2873
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2966..3095
                     /label=AAV2 ITR
     primer_bind     complement(3114..3131)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3114..3130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3123..3145)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3272..3727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3359..3378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3569..3590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3753..3857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3858..3926,3927..4718)
                     /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(4076..4095)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4889..5477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5378..5397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5765..5786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5801..5806,5807..5824,5825..5831)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5839..5855
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5844..5866
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5863..5879
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5863..5879
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacca gggtaatggg
      181 gatcctctag aactatagct agaattcgcc cttaagctag caggttaatt tttaaaaagc
      241 agtcaaaagt ccaagtggcc cttggcagca tttactctct ctgtttgctc tggttaataa
      301 tctcaggagc acaaacattc cagatccagg ttaattttta aaaagcagtc aaaagtccaa
      361 gtggcccttg gcagcattta ctctctctgt ttgctctggt taataatctc aggagcacaa
      421 acattccaga tccggcgcgc cagggctgga agctaccttt gacatcattt cctctgcgaa
      481 tgcatgtata atttctacag aacctattag aaaggatcac ccagcctctg cttttgtaca
      541 actttccctt aaaaaactgc caattccact gctgtttggc ccaatagtga gaactttttc
      601 ctgctgcctc ttggtgcttt tgcctatggc ccctattctg cctgctgaag acactcttgc
      661 cagcatggac ttaaacccct ccagctctga caatcctctt tctcttttgt tttacatgaa
      721 gggtctggca gccaaagcaa tcactcaaag ttcaaacctt atcatttttt gctttgttcc
      781 tcttggcctt ggttttgtac atcagctttg aaaataccat cccagggtta atgctggggt
      841 taatttataa ctaagagtgc tctagttttg caatacagga catgctataa aaatggaaag
      901 atgttgcttt ctgagagaca gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      961 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc acatggaaga cgccaaaaac
     1021 ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg
     1081 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
     1141 atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct
     1201 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
     1261 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
     1321 gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg
     1381 ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc
     1441 caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
     1501 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc
     1561 gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa
     1621 ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt
     1681 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     1741 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     1801 agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg
     1861 ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta
     1921 tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg
     1981 gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca
     2041 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     2101 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     2161 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     2221 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     2281 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     2341 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     2401 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     2461 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     2521 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     2581 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     2641 aagatcgccg tgtaattcta gattccgaga tatcggtaat gggcccgcgt ggtacctcta
     2701 gagtcgaccc gggcggcctc gaggacgggg tgaactacgc ctgaggatcc gatctttttc
     2761 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2821 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2881 gcaattcgtt gatctgaatt tcgaccaccc ataataccca ttaccctggt agataagtag
     2941 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3001 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3061 gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc
     3121 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3181 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3241 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3301 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3361 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3421 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3481 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3541 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3601 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3661 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3721 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3781 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3841 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3901 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3961 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4021 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4081 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4141 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4201 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4261 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4321 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4381 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4441 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4501 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4561 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4621 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4681 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4741 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4801 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4861 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4921 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4981 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5041 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5101 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5161 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5221 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5281 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5341 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5401 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5461 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5521 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5581 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5641 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5701 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     5761 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     5821 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     5881 atgattacgc cagatttaat taaggcctta attag
//
