LOCUS       sequence-43848-e        4734 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 4734)
  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 4734)
  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: pAAVscCBPIGpluc
FEATURES             Location/Qualifiers
     source          1..4734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          797..893
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             974..1531
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     primer_bind     complement(1547..1564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1553..1664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1784..1913
                     /label=AAV2 ITR
     primer_bind     complement(1932..1949)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1932..1948)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1941..1963)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2090..2545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2177..2196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2387..2408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2571..2675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2676..2744,2745..3536)
                     /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(2894..2913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3707..4295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4196..4215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4583..4604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4619..4624,4625..4642,4643..4649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4657..4673
                     /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     4662..4684
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4681..4697
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4681..4697
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtgtagcc atgctctagg
      121 aagatcaatt cggtacaatt cacgcgtcga cattgattat tgactctggt cgttacataa
      181 cttacggtaa atggcccgcc tggctgaccg cccaacgacc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      421 gggactttcc tacttggcag tacatctact cgaggccacg ttctgcttca ctctccccat
      481 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      541 gatgggggcg gggggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
      601 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      661 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      721 gggcgggagc gggatcagcc accgcggtgg cggccctaga gtcgatcgag gaactgaaaa
      781 accagaaagt taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg
      841 tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta
      901 cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccgcggccga tccaccggtc
      961 gccaccatct agcatgggag tcaaagttct gtttgccctg atctgcatcg ctgtggccga
     1021 ggccaagccc accgagaaca acgaagactt caacatcgtg gccgtggcca gcaacttcgc
     1081 gaccacggat ctcgatgctg accgcgggaa gttgcccggc aagaagctgc cgctggaggt
     1141 gctcaaagag atggaagcca atgcccggaa agctggctgc accaggggct gtctgatctg
     1201 cctgtcccac atcaagtgca cgcccaagat gaagaagttc atcccaggac gctgccacac
     1261 ctacgaaggc gacaaagagt ccgcacaggg cggcataggc gaggcgatcg tcgacattcc
     1321 tgagattcct gggttcaagg acttggagcc catggagcag ttcatcgcac aggtcgatct
     1381 gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt gttctgacct
     1441 gctcaagaag tggctgccgc aacgctgtgc gacctttgcc agcaagatcc agggccaggt
     1501 ggacaagatc aagggggccg gtggtgacta gctcgacgct gatcagcctc gactgtgcct
     1561 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1621 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1681 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1741 aattaggtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     1801 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     1861 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat
     1921 taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1981 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2041 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     2101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2281 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2401 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2521 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2581 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2641 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2701 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2761 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2821 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2881 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2941 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3001 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3061 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3121 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3181 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3241 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3301 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3361 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3421 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3481 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3541 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3601 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3661 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4321 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4381 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4441 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4501 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4561 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4621 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4681 caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat tagg
//
