LOCUS       sequence-133591-        4455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBT264-[miR-488].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4455)
  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 4455)
  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-34
FEATURES             Location/Qualifiers
     source          1..4455
                     /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(2270..2288)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2309..2325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2309..2325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2322..2344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2333..2349
                     /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(2357..2363,2364..2381,2382..2387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2402..2423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2540..2557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2711..3299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2791..2810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3470..4261,4262..4330))
                     /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     4093..4112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4331..4435)
                     /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 aacgcgaaga attccgttgc ggatcccctc aactacactg cccccaatcc
     1321 ttgcctagct caaacccaag gccaatgtgg tagaggaagc aggggtacat atagtatcca
     1381 tcttttcaca tacaacacac tttattaact ttgtttgtct gaacagcaaa atatgactcc
     1441 tgagtcttcc agaagaaatt gagagtcatc tgaccaagaa atagcctttc aaacaatttg
     1501 gaaacttgtt tgagagtgcc attatctggg agagatgatt ctctgaaatt ttccactcta
     1561 ggctcttctt tcttccttag gaggaaacac cagttgtggt ccaaaaattc ttcttgcaga
     1621 aactggtttt gttccataga gcccacaacc caacccaaca cctcttcgcc ttcccccgcc
     1681 tccctcactc ccagatcctt taccaccttc tgccacctct atcattccta cctctactac
     1741 gccagatctt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     1801 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     1861 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     1921 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     1981 gggggctgtc cccgtgcact cagcctaaag ctttttcccc gtatcccccc aggtgtctgc
     2041 aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg
     2101 cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg
     2161 gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg
     2221 ggctttgggg gggggctgtc cccatttaaa tcagctccag cttttgttcc ctttagtgag
     2281 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3121 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4441 ttccccgaaa agtgc
//
