LOCUS       sequence-121066-        5182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  8x Target 2 0gap with SV40 Ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5182)
  AUTHORS   Slomovic S, Collins JJ
  TITLE     DNA sense-and-respond protein modules for mammalian cells.
  JOURNAL   Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
  PUBMED    26389572
REFERENCE   2  (bases 1 to 5182)
  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 
            Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585."
COMMENT     Sequence Label: pBW121-SS-321
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1845..1864)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1910..1965
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1964..1983)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2324..2340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2324..2340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2337..2359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2348..2364
                     /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(2372..2378,2379..2396,2397..2402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2417..2438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2873..3007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3027..3161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3064..3083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3118..3137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3261..3278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3432..4020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3512..3531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4191..4982,4983..5051))
                     /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     4814..4833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5052..5156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgattaat tcgagcgaac
     1741 gcgtagccaa aggcaacagg cggccgcact cctcaggtgc aggctgccta tcagaaggtg
     1801 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     1861 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     1921 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     1981 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     2041 gccatatgct ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca
     2101 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     2161 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     2221 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     2281 atgaagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     2341 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2401 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2461 ctttccagtc gggaaacctg tcgtgccagc ggatctagag tgaggagccg aagatgggga
     2521 cgtcgtgtag gggtggatct agagtgagga gccgaagatg gggacgtcgt gtaggggtgg
     2581 atctagagtg aggagccgaa gatggggacg tcgttagggg tggatctaga gtgaggagcc
     2641 gaagatgggg acgtcgtgta ggggtggatc tagagtgagg agccgaagat ggggacgtcg
     2701 tgtaggggtg gatctagagt gaggagccga agatggggac gtcgtgtagg ggtggatcta
     2761 gagtgaggag ccgaagatgg ggacgtcgtt aggggtggat ctagagtgag gagccgaaga
     2821 tggggacgtc gtgtaggggt ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     2881 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2941 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3001 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     3061 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3121 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgcgaaatt aagcgaaatt
     3181 aactcgagag atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3601 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     3661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3841 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4201 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4261 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4321 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4381 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4441 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4501 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4561 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4621 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4681 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4741 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4801 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4861 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4921 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4981 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5041 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5161 tttccccgaa aagtgccacc tg
//
