LOCUS       sequence-133590-        4645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBT264-[miR-429].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4645)
  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 4645)
  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-32
FEATURES             Location/Qualifiers
     source          1..4645
                     /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(2460..2478)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2499..2515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2499..2515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2512..2534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2523..2539
                     /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(2547..2553,2554..2571,2572..2577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2592..2613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2730..2747)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2901..3489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2981..3000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3660..4451,4452..4520))
                     /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     4283..4302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4521..4625)
                     /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 ctgaatgaga attccgttgc ggatccctcc gggtatctgt gactgtgacc
     1321 ccaggggcca ggctgatagc acggggtcca cagcatctgc ccggggacta cagctcgagg
     1381 gccccaggag gccgggtggg cacaggccat ggcgagggtg gggcacaaga gccccagacc
     1441 ccggcggctt tgcactgatg ggctgcggat gggcacaggc catagtgagg ggggcatgag
     1501 agccccagac cgggcggctt tgcactgatg agctgcaggg caggcagctc tctcctctaa
     1561 cggtgatcag gcagcggatg gacggtttta ccagacagta ttagacagag ggccaggtct
     1621 aaccatgtct ggtaagacgc ccatcggccg gcggtggtgt gggtggcctc cgggtcctgg
     1681 gctggtgtct gcggggcctg ngaagggaag gccccggggg cggctcctgc cacctcctgc
     1741 agctgccctg atccctcctg ggtctgcaga acgagccccc gaggcacttc cctgacagcc
     1801 cccatcccag cacctggtcc cagcccagcc tggctgtgtg ggggtacccg ggagccacct
     1861 ggtcacccag gccctgctct tggcaaaggt tccctgcagc ccctcgctgg cttcctgaag
     1921 ctcggtcggg ttaagatctt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc
     1981 aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg
     2041 cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg
     2101 gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg
     2161 ggctttgggg gggggctgtc cccgtgcact cagcctaaag ctttttcccc gtatcccccc
     2221 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     2281 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     2341 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     2401 catccctggg ggctttgggg gggggctgtc cccatttaaa tcagctccag cttttgttcc
     2461 ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2521 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2581 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2641 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2701 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2761 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2821 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2881 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2941 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3001 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3061 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3121 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3181 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3241 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3301 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3361 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3421 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3481 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3541 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3601 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3661 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3721 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3781 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3841 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3901 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3961 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4021 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4081 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4141 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4201 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4261 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4321 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4381 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4441 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4501 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4561 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4621 ccgcgcacat ttccccgaaa agtgc
//
