LOCUS       sequence-177012-        5395 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PV7 for 3-input, 2-output Full Adder.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5395)
  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 5395)
  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: pBW810_pCAG-PV7-loxN-F14-mCherry-BGHpA
FEATURES             Location/Qualifiers
     source          1..5395
                     /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    complement(668..701)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      746..755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             752..1462
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(897..915)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1165..1184
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1463..1687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1933..1952)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1998..2053
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2052..2071)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2414..2430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2414..2430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2427..2449)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2438..2454
                     /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(2462..2468,2469..2486,2487..2492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2507..2528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2613..2747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2767..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2804..2823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2858..2877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2969..2986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3140..3728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3899..4690,4691..4759))
                     /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     4522..4541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4760..4864)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4895..5274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
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 aagacgctgg ctctgacatt tccgctactg aactactcgg
      661 cacgcgtata acttcgtata gtatacctta tacgaagtta tcgaagttcc tattctatca
      721 gaagtatagg aacttcaccg gtgccgccac catggtgagc aagggcgagg aggataacat
      781 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      841 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
      901 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      961 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1021 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1081 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1141 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1201 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1261 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1321 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1381 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1441 catggacgag ctgtacaagt aactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1501 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1561 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1621 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1681 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1741 tgtagcggaa ttcggtaccc ctcgtctcaa ccaaagcaat caacccatca accacctggg
     1801 cggccgccca ggatacatag attaccacaa ctccgagccc ttccaccgct agcgcactcc
     1861 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     1921 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     1981 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2041 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     2101 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     2161 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     2221 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     2281 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     2341 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     2401 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2461 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2521 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2581 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccagttccgc
     2641 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2701 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2761 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2821 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2881 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2941 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4861 cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa
     4921 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4981 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5041 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5101 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5161 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5221 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     5281 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     5341 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg ggggg
//
