LOCUS       sequence-177024-        4317 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Simple PV8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4317)
  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 4317)
  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: pBW754_pCAG-PV8
FEATURES             Location/Qualifiers
     source          1..4317
                     /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"
     primer_bind     complement(855..874)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    920..975
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(974..993)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1336..1352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1336..1352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1349..1371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1360..1376
                     /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(1384..1390,1391..1408,1409..1414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1429..1450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1535..1669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1689..1823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1726..1745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1780..1799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1891..1908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2062..2650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2142..2161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2821..3612,3613..3681))
                     /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     3444..3463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3682..3786)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3817..4196
                     /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 ctcgtctcaa ccaaagcaat caacccatca accacctggg
      661 cacgcgtaag tctacggtac cgttccttat catctggcga atcggaccca caagagcact
      721 ggcggccgcc caggatacat agattaccac aactccgagc ccttccaccg ctagcgcact
      781 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
      841 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
      901 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
      961 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     1021 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     1081 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     1141 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     1201 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     1261 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     1321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1501 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccagttcc
     1561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     1681 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1741 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1801 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     1861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2581 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2641 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2701 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2761 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2821 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2881 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2941 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3001 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3061 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3121 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3181 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3241 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3301 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3361 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3421 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3481 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3541 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3601 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3661 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3781 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
     3841 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3901 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3961 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4021 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4081 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4141 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     4201 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     4261 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggg
//
