LOCUS       sequence-177040-        4267 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCAG destination vector (U5-UX).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4267)
  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 4267)
  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: pBW755_pCAG-pDest-U5-UX
FEATURES             Location/Qualifiers
     source          1..4267
                     /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(807..826)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    872..927
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(926..945)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1288..1304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1288..1304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1301..1323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1312..1328
                     /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(1336..1342,1343..1360,1361..1366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1381..1402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1487..1621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1641..1775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1678..1697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1732..1751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1843..1860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2773..3564,3565..3633))
                     /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     3396..3415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3634..3738)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3769..4148
                     /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 aggcgcgccg agccaactcc ctttacaacc tcactcaagt ccgttagagg
      661 cggccgcaag tctacgaatt cccaggatac atagattacc acaactccga gcccttccac
      721 cgctagcgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
      781 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
      841 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
      901 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
      961 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     1021 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     1081 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     1141 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     1201 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     1261 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1441 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1501 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1561 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     1621 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     1681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1741 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     1801 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1861 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3721 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat
     3781 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3841 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3901 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3961 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4021 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4081 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4141 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     4201 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     4261 ggggggg
//
