LOCUS       sequence-43860-e        6044 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 6044)
  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 6044)
  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 miR122sponge
FEATURES             Location/Qualifiers
     source          1..6044
                     /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(2882..2901)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2947..3002
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3095..3224
                     /label=AAV2 ITR
     primer_bind     complement(3243..3260)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3243..3259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3252..3274)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3401..3856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3488..3507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3698..3719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3882..3986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3987..4055,4056..4847)
                     /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(4205..4224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5018..5606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5507..5526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5894..5915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5930..5935,5936..5953,5954..5960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5968..5984
                     /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     5973..5995
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5992..6008
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5992..6008
                     /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 gacaaacacc atacaacact ccacaaacac catacaacac
     2701 tccacaaaca ccatacaaca ctccacaaac accatacaac actccacaaa caccatacaa
     2761 cactccacaa acaccataca acactccaca aacaccatac aacactccag ggcccgcgtg
     2821 gtacctctag agtcgacccg ggcggcctcg aggacggggt gaactacgcc tgaggatccg
     2881 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     2941 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3001 cactcggaag caattcgttg atctgaattt cgaccaccca taatacccat taccctggta
     3061 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc
     3121 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3181 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat
     3241 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3301 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3361 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3421 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3481 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3541 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3601 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3661 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3721 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3781 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3841 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3961 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4021 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4081 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4141 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4201 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4261 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4321 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4381 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4441 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4501 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4561 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4621 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4681 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4741 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4801 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4861 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5161 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5641 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5701 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5761 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5821 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5881 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5941 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6001 gctatgacca tgattacgcc agatttaatt aaggccttaa ttag
//
