LOCUS       sequence-43845-e        7647 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 7647)
  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 7647)
  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: pAAVf-EnhCB-lacZnls
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..142
                     /label=AAV2 ITR
     intron          830..926
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        990..1037
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        1029..1047
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1051..1060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1288..1308
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4308..4325)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4314..4538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4845..4862)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4845..4861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4854..4876)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5003..5458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5090..5109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5300..5321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5484..5588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5589..5657,5658..6449)
                     /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(5807..5826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6620..7208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7109..7128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7496..7517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7532..7537,7538..7555,7556..7562)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7570..7586
                     /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     7575..7597
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7594..7610
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7594..7610
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gccttaatta ggctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg
      181 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      241 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      301 caatgggtgg agtattttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      361 gccaagtacg ccccctattg ccgtcaatga cggtaaatgg cccgcctggc attatgccca
      421 gtacatgacc ttatgggact ttcctacttg gcagtacatc tactcgaggc cacgttctgc
      481 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
      541 ttattttgtg cagcgatggg ggcggggggg gggggcgcgc gccaggcggg gcggggcggg
      601 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
      661 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
      721 cgcggcgggc gggagcggga tcagaatgat ctgatatcat cgatgaattc gagctcggta
      781 cccggggatc ctctagagtc gaggaactga aaaaccagaa agttaactgg taagtttagt
      841 ctttttgtct tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc
      901 tcagtggatg ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc
      961 tgcggaattg tacccgcggc caaacagtag ttgacaatta atcatcggca tagtatatcg
     1021 gcatagtata atacgactca ctataggagg gccaccatgg accctgttgt gctgcaaagg
     1081 agagactggg agaaccctgg agtgacccag ctcaacagac tggctgccca ccctcccttt
     1141 gcctcttgga ggaactctga ggaagccagg acagacaggc ccagccagca gctcaggtct
     1201 ctcaatggag agtggaggtt tgcctggttc cctgcccctg aagctgtgcc tgagtcttgg
     1261 ctggagtgtg acctcccaga ggcagttccc aagaagaaga ggaaagttga ggctgacact
     1321 gttgtggtgc caagcaactg gcagatgcat ggctatgatg cccccatcta caccaatgtc
     1381 acctacccca tcactgtgaa cccccctttt gtgcccactg agaaccccac tggctgctac
     1441 agcctgacct tcaatgttga tgagagctgg ctgcaagaag gccagaccag gatcatcttt
     1501 gatggagtca actctgcctt ccacctctgg tgcaatggca ggtgggttgg ctatggccaa
     1561 gacagcaggc tgccctctga gtttgacctc tctgccttcc tcagagctgg agagaacagg
     1621 ctggctgtca tggtgctcag gtggtctgat ggcagctacc tggaagacca agacatgtgg
     1681 aggatgtctg gcatcttcag ggatgtgagc ctgctgcaca agcccaccac ccagatttct
     1741 gacttccatg ttgccaccag gttcaatgat gacttcagca gagctgtgct ggaggctgag
     1801 gtgcagatgt gtggagaact cagagactac ctgagagtca cagtgagcct ctggcaaggt
     1861 gagacccagg tggcctctgg cacagccccc tttggaggag agatcattga tgagagagga
     1921 ggctatgctg acagagtcac cctgaggctc aatgtggaga accccaagct gtggtctgct
     1981 gagatcccca acctctacag ggctgttgtg gagctgcaca ctgctgatgg caccctgatt
     2041 gaagctgaag cctgtgatgt tggattcaga gaagtcagga ttgagaatgg cctgctgctg
     2101 ctcaatggca agcctctgct catcagggga gtcaacaggc atgagcacca ccctctgcat
     2161 ggacaagtga tggatgaaca gacaatggtg caagatatcc tgctaatgaa gcagaacaac
     2221 ttcaatgctg tcaggtgctc tcactacccc aaccaccctc tctggtacac cctgtgtgac
     2281 aggtatggcc tgtatgttgt tgatgaagcc aacattgaga cacatggcat ggtgcccatg
     2341 aacaggctca cagatgaccc caggtggctg cctgccatgt ctgagagagt gaccaggatg
     2401 gtgcagagag acaggaacca cccctctgtg atcatctggt ctctgggcaa tgagtctgga
     2461 catggagcca accatgatgc tctctacagg tggatcaagt ctgttgaccc cagcagacct
     2521 gtgcagtatg aaggaggtgg agcagacacc acagccacag acatcatctg ccccatgtat
     2581 gccagggttg atgaggacca gcccttccct gctgtgccca agtggagcat caagaagtgg
     2641 ctctctctgc ctggagagac cagacctctg atcctgtgtg aatatgcaca tgcaatgggc
     2701 aactctctgg gaggctttgc caagtactgg caagccttca gacagtaccc caggctgcaa
     2761 ggaggatttg tgtgggactg ggtggaccaa tctctcatca agtatgatga gaatggcaac
     2821 ccctggtctg cctatggagg agactttggt gacaccccca atgacaggca gttctgcatg
     2881 aatggcctgg tctttgcaga caggacccct caccctgccc tcacagaggc caagcaccag
     2941 caacagttct tccagttcag gctgtctgga cagaccattg aggtgacatc tgagtacctc
     3001 ttcaggcact ctgacaatga gctcctgcac tggatggtgg ccctggatgg caagcctctg
     3061 gcttctggtg aggtgcctct ggatgtggcc cctcaaggaa agcagctgat tgaactgcct
     3121 gagctgcctc agccagagtc tgctggacaa ctgtggctaa cagtgagggt ggttcagccc
     3181 aatgcaacag cttggtctga ggcaggccac atctctgcat ggcagcagtg gaggctggct
     3241 gagaacctct ctgtgaccct gcctgctgcc tctcatgcca tccctcacct gacaacatct
     3301 gaaatggact tctgcattga gctgggcaac aagagatggc agttcaacag gcagtctggc
     3361 ttcctgtctc agatgtggat tggagacaag aagcagctcc tcacccctct cagggaccaa
     3421 ttcaccaggg ctcctctgga caatgacatt ggagtgtctg aggccaccag gattgaccca
     3481 aatgcttggg tggagaggtg gaaggctgct ggacactacc aggctgaggc tgccctgctc
     3541 cagtgcacag cagacaccct ggctgatgct gttctgatca ccacagccca tgcttggcag
     3601 caccaaggca agaccctgtt catcagcaga aagacctaca ggattgatgg ctctggacag
     3661 atggcaatca cagtggatgt ggaggttgcc tctgacacac ctcaccctgc aaggattggc
     3721 ctgaactgtc aactggcaca ggtggctgag agggtgaact ggctgggctt aggccctcag
     3781 gagaactacc ctgacaggct gacagctgcc tgctttgaca ggtgggacct gcctctgtct
     3841 gacatgtaca ccccttatgt gttcccttct gagaatggcc tgaggtgtgg caccagggag
     3901 ctgaactatg gtcctcacca gtggagggga gacttccagt tcaacatctc caggtactct
     3961 cagcaacagc tcatggaaac ctctcacagg cacctgctcc atgcagagga gggaacctgg
     4021 ctgaacattg atggcttcca catgggcatt ggaggagatg actcttggtc tccttctgtg
     4081 tctgctgagt tccagttatc tgctggcagg taccactatc agctggtgtg gtgccagaag
     4141 taaacctgag ctagcggatc catatatagg gcccgggtta taattacctc aggtcgacgt
     4201 cccatggcca ttcgaaacaa acaccattgt cacactccaa caaacaccat tgtcacactc
     4261 caacaaacac cattgtcaca ctccattcga attaaaccgc tgatcagcct cgactgtgcc
     4321 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4381 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4441 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4501 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4561 cagatctgca gatctgaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4621 ggttttttcg gatcctctag agtcgaccgt agataagtag catggcgggt taatcattaa
     4681 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4741 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4801 cgagcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta
     4861 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4921 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4981 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5041 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5101 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5161 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5221 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5281 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5341 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5401 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5461 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5521 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5581 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5701 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5761 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5881 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6001 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6121 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6181 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6421 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6481 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6541 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6601 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6661 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6721 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6781 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6841 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6901 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6961 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7021 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7081 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7141 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7201 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7261 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7321 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     7381 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     7441 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7501 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     7561 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     7621 ccagatttaa ttaaggcctt aattagg
//
