LOCUS       sequence-133548-        4501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBT264-[miR-132].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4501)
  AUTHORS   Wong AS, Choi GC, Cheng AA, Purcell O, Lu TK
  TITLE     Massively parallel high-order combinatorial genetics in human cells.
  JOURNAL   Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub 
            2015 Aug 17.
  PUBMED    26280411
REFERENCE   2  (bases 1 to 4501)
  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/nbt.3326"; 
            journalName: "Nat Biotechnol"; date: "2015-09"; volume: "33"; issue:
            "9"; pages: "952-61"
COMMENT     Sequence Label: pAWSV-11
FEATURES             Location/Qualifiers
     source          1..4501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1175..1256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1200..1219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2316..2334)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2355..2371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2355..2371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2368..2390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2379..2395
                     /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)."
     promoter        complement(join(2403..2409,2410..2427,2428..2433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2448..2469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2757..3345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2837..2856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3516..4307,4308..4376))
                     /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     4139..4158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4377..4481)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtacagg
      661 cgcgcctcag cctaaagctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc
      721 agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc
      781 cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg
      841 ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg
      901 ggctgtcccc gtgcactcag cctaaagctt tttccccgta tccccccagg tgtctgcagg
      961 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc
     1021 gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag
     1081 ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc
     1141 tttggggggg ggctgtcccc actagtgatt cgaggcagtg aaaaaaatgc tttatttgtg
     1201 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1261 acaacaattg taggacgcga attccgttgc ggatccgagc cctggctggg ataccttggc
     1321 cgggaggaca ggacggcctc aggctggccg ccagacgcac tccgcaggcc gcgcgcgctc
     1381 gagcctcggt ggactcagcc gccggccccg ggaggccgcc tccctgcgca gcacctcctg
     1441 ccacccgcgc gggcacgtgg gatcttgact cgccgccccc gcctccttct gctccgcgtc
     1501 cccagcccgc ggctcggggc gcggcgtggc gcggcgcgtg ggcgtgctgc ggggcgacca
     1561 tggctgtaga ctgttacctc cagttcccac agtaacaatc gaaagccacg gttgccctgg
     1621 agacgcgggg gcggggcgcg cggacggcgc cggcgcgggg cgggctgacg tcagggccgc
     1681 gggggggcgg gcggggcggc gggggctgag gagcgcgcgt gggccaccgc ccctcgcagc
     1741 ccctagggac ccggacgggg cggcccgatg gcggcgcctg cgttgatcaa gatcttcagc
     1801 ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     1861 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     1921 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     1981 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccccg
     2041 tgcactcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca
     2101 gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc
     2161 gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc
     2221 tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg
     2281 gctgtcccca tttaaatcag ctccagcttt tgttcccttt agtgagggtt aatttcgagc
     2341 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3181 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4501 c
//
