LOCUS       sequence-176802-        4052 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PV1 for 2-input, 2-output circuit library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4052)
  AUTHORS   Weinberg BH, Pham NT, Caraballo LD, Lozanoski T, Engel A, Bhatia S, 
            Wong WW
  TITLE     Large-scale design of robust genetic circuits with multiple inputs 
            and outputs for mammalian cells.
  JOURNAL   Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805.
  PUBMED    28346402
REFERENCE   2  (bases 1 to 4052)
  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"; journalName: "Nat 
            Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805."
COMMENT     Sequence Label: pBW705_pCAG-PV1-FRT-lox2272-STOP
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    668..701
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(703..736)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             785..1579
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(839..858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1449..1468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1759..1893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2087..2106)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2152..2207
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2206..2225)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2568..2584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2568..2584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2581..2603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2592..2608
                     /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(2616..2622,2623..2640,2641..2646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2661..2682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2767..2901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2921..3055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga aggcgcgccc attactcgca tccattctca ggctgtctcg tctcgtctcg
      661 cacgcgtgaa gttcctattc tctagaaagt ataggaactt cgataacttc gtataggata
      721 ctttatacga agttataccg gtcggatttg atctgatcaa gagacaggat gaggatcgtt
      781 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      841 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      901 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      961 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1021 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1081 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1141 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1201 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1261 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1321 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1381 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1441 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1501 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1561 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1621 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     1681 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1741 ttcgcccacc ccatcgataa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1801 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1861 aaactcatca atgtatctta tcatgtctgg atcaaatccg agaattcggt accgctggga
     1921 gttcgtagac ggaaacaaac gcagaatcca agcgcggccg cccaggatac atagattacc
     1981 acaactccga gcccttccac cgctagcgca ctcctcaggt gcaggctgcc tatcagaagg
     2041 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     2101 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     2161 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     2221 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     2281 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     2341 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     2401 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     2461 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     2521 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     2581 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2641 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2701 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     2761 ccatagtccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2821 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2881 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4021 taaagtatat atgagtaaac ttggtctgac ag
//
