LOCUS       sequence-125008-        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  8x Target 1 4gap minus SV40 Ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  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 4900)
  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-310
FEATURES             Location/Qualifiers
     source          1..4900
                     /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."
     primer_bind     complement(2979..2996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3150..3738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3230..3249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3909..4700,4701..4769))
                     /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     4532..4551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4770..4874)
                     /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 ggatcttgaa gaagctgcag gaggtgtcag
     2521 ctggagggga agtggtccgg atcttgaaga agctgcagga ggtgtcagct ggaggggaag
     2581 tggtccggat cttgaagaag ctgcaggagg tgtcagctgg aggggaagtg gtccggatct
     2641 tgaagaagct gcaggaggtg tcagctggag gggaagtggt ccggatcttg aagaagctgc
     2701 aggaggtgtc agctggaggg gaagtggtcc ggatcttgaa gaagctgcag gaggtgtcag
     2761 ctggagggga agtggtccgg atcttgaaga agctgcagga ggtgtcagct ggaggggaag
     2821 tggtccggat cttgaagaag ctgcaggagg tgtcagctgg aggggaagtg gtccggatcc
     2881 gcgaaattaa gcgaaattaa ctcgagagat ccgctgcatt aatgaatcgg ccaacgcgcg
     2941 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     3361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3901 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3961 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4021 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4081 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4141 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4201 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4261 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4321 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4381 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4441 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4501 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4561 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4621 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4681 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4741 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4801 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
