LOCUS       sequence-177141-        4166 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCAG destination vector (U1-UX).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4166)
  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 4166)
  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: pBW700_pCAG-pDest-U1-UX
FEATURES             Location/Qualifiers
     source          1..4166
                     /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"
     primer_bind     complement(687..706)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    752..807
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(806..825)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1168..1184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1168..1184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1181..1203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1192..1208
                     /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(1216..1222,1223..1240,1241..1246))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1261..1282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1367..1501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1521..1655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1558..1577)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1612..1631
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1723..1740)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1894..2482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1974..1993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2653..3444,3445..3513))
                     /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     3276..3295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3514..3618)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3649..4028
                     /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 attactcgca tccattctca ggctgtctcg tctcgtctcg
      541 cggccgcaag tctacgaatt cccaggatac atagattacc acaactccga gcccttccac
      601 cgctagcgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
      661 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
      721 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
      781 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
      841 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
      901 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
      961 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     1021 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     1081 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     1141 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1201 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1261 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1321 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1441 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     1501 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     1561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1621 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     1681 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1741 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1801 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2401 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2761 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2941 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3481 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3541 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3601 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat
     3661 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3721 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3781 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3841 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3901 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3961 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4021 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     4081 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     4141 gggggggggc gcgcgccagg cggggc
//
