LOCUS       sequence-177132-        4105 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCAG promoter driving no CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4105)
  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 4105)
  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: pCAG-FALSE
FEATURES             Location/Qualifiers
     source          1..4105
                     /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(720..739)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    785..840
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(839..858)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1201..1217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1201..1217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1214..1236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1225..1241
                     /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(1249..1255,1256..1273,1274..1279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1294..1315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1400..1534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1554..1688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1591..1610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1645..1664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1756..1773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1927..2515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2007..2026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2686..3477,3478..3546))
                     /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     3309..3328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3547..3651)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3682..4061
                     /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 acgcgtcgac ggaccggttt cacggcggcc gcactcctca ggtgcaggct
      661 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
      721 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
      781 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
      841 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
      901 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
      961 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     1021 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     1081 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     1141 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     1201 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     1381 aattagtcag caaccatagt cccgccccta actccgccca gttccgccca ttctccgccc
     1441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     1561 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1621 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1681 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2821 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3661 ccgaaaagtg ccacctgggt cgacattgat tattgactag ttattaatag taatcaatta
     3721 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     3781 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     3841 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     3901 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     3961 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4021 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
     4081 tctgcttcac tctccccatc tcccc
//
