LOCUS       sequence-43846-e        7601 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 7601)
  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 7601)
  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 mir-122 1xBS
FEATURES             Location/Qualifiers
     source          1..7601
                     /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(4262..4279)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4268..4492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4799..4816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4799..4815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4808..4830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4957..5412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5044..5063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5254..5275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5438..5542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5543..5611,5612..6403)
                     /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(5761..5780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6574..7162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7063..7082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7450..7471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7486..7491,7492..7509,7510..7516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7524..7540
                     /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     7529..7551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7548..7564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7548..7564
                     /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 cacactccat tcgaattaaa ccgctgatca
     4261 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4321 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4381 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4441 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4501 gcggaaagaa ccagcagatc tgcagatctg aattgcattc attttatgtt tcaggttcag
     4561 ggggaggtgt gggaggtttt ttcggatcct ctagagtcga ccgtagataa gtagcatggc
     4621 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4681 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4741 gcggcctcag tgagcgagca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac
     4801 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     4861 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     4921 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     4981 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5041 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5101 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5161 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5221 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5281 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5341 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5401 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6721 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7321 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7381 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7441 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7501 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7561 tgaccatgat tacgccagat ttaattaagg ccttaattag g
//
