LOCUS       sequence-177031-        5236 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PV4 for 3-input, 2-output Full Adder.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5236)
  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 5236)
  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: pBW807_pCAG-PV4-loxP-mCherry-BGHpA
FEATURES             Location/Qualifiers
     source          1..5236
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     346..368
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     430..449
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(548..581)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      591..600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             597..1307
                     /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(742..760)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1010..1029
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1308..1532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1778..1797)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1843..1898
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1897..1916)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2259..2275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2259..2275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2272..2294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2283..2299
                     /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(2307..2313,2314..2331,2332..2337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2352..2373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2458..2592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2612..2746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2649..2668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2703..2722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2814..2831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2985..3573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3065..3084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3744..4535,4536..4604))
                     /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     4367..4386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4605..4709)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4740..5119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
       61 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      121 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      181 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      241 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      301 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      361 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      421 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      481 attcgagcga aggcgcgccc tgacctcctg ccagcaatag taagacaaca cgcaaagtcg
      541 cacgcgtata acttcgtata gcatacatta tacgaagtta taccggtgcc gccaccatgg
      601 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
      661 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      721 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
      781 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
      841 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
      901 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
      961 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1021 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1081 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1141 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1201 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     1261 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaactg tgccttctag
     1321 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1381 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1441 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1501 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1561 ctctaggggg tatccccacg cgccctgtag cggaattcgg taccgagcca actcccttta
     1621 caacctcact caagtccgtt agaggcggcc gcccaggata catagattac cacaactccg
     1681 agcccttcca ccgctagcgc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     1741 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     1801 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     1861 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     1921 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     1981 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     2041 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     2101 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     2161 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     2221 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     2281 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2341 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2401 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     2461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2521 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2581 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     2641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2701 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     2761 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2821 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2881 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2941 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3001 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3061 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3121 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3181 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3241 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3301 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3361 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3421 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3481 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3541 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3601 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3661 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3721 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3781 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3841 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3901 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3961 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4021 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4081 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4141 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4201 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4261 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4321 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4381 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4441 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4501 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4561 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4621 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4681 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg
     4741 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     4801 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4861 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4921 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4981 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5041 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     5101 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     5161 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     5221 gggggcgggg gggggg
//
