LOCUS       sequence-121029-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CIS-V2m with N-FLAG and C-HIS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  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 5863)
  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-279
FEATURES             Location/Qualifiers
     source          1..5863
                     /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"
     CDS             1748..1771
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1772..1792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2794..2805
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2918..3067
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             3086..3103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3192..3211)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3257..3312
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3311..3330)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3671..3687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3671..3687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3684..3706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3695..3711
                     /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(3719..3725,3726..3743,3744..3749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3764..3785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3942..3959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4113..4701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4193..4212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4872..5663,5664..5732))
                     /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     5495..5514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5733..5837)
                     /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 gcgtatggac tacaaggacg acgatgacaa gcccaagaaa aagcgcaagg tctctagacc
     1801 cggggagcgc cccttccagt gtcgcatttg catgcggaac ttttcggata aaactaaatt
     1861 gagagttcat acccgtactc ataccggtga aaaaccgttt cagtgtcgga tctgtatgcg
     1921 aaatttctcc gttagacata atttgactag acatctacgt acgcacaccg gcgagaagcc
     1981 attccaatgc cgaatatgca tgcgcaactt cagtcaatct acttctttgc aaagacacct
     2041 aaaaacccac ctgagaggat ccggcggagt gctggagaag ggcgcctttg ccaagggtac
     2101 caatgtttta atggcggatg ggtctattga atgtattgaa aacattgagg ttggtaataa
     2161 ggtcatgggt aaagatggca gacctcgtga ggtaattaaa ttgcccagag gaagagaaac
     2221 tatgtacagc gtcgtgcaga aaagtcagca cagagcccac aaaagtgact caagtcgtga
     2281 agtgccagaa ttactcaagt ttacgtgtaa tgcgacccat gagttggttg ttagaacacc
     2341 tcgtagtgtc cgccgtttgt ctcgtaccat taagggtgtc gaatattttg aagttattac
     2401 ttttgagatg ggccaaaaga aagcccccga cggtagaatt gttgagcttg tcaaggaagt
     2461 ttcaaagagc tacccaatat ctgaggggcc tgagagagcc aacgaattag tagaatccta
     2521 tagaaaggct tcaaataaag cttattttga gtggactatt gaggccagag atctttctct
     2581 gttgggttcc catgttcgta aagctaccta ccagacttac gctccaattc tttatggcct
     2641 gaggggatct gggggcggat caggtggcgg atcaggaggc ggatcagttt tgcttaacgt
     2701 tctttcgaag tgtgccggct ctaaaaaatt caggcctgct cccgccgctg cttttgcacg
     2761 tgagtgccgc ggattttatt tcgagttaca agaattgaag gaagacgatt attatgggat
     2821 tactttatct gatgattctg atcatcagtt tttgcttgcc aaccaggttg tcgtccatgc
     2881 ttgcaccatg accgagaagg gcagcggagg gcgcgccgac gcgctggacg atttcgatct
     2941 cgacatgctg ggttctgatg ccctcgatga ctttgacctg gatatgttgg gaagcgacgc
     3001 attggatgac tttgatctgg acatgctcgg ctccgatgct ctggacgatt tcgatctcga
     3061 tatgttaatt aattgtggat ctggccacca tcaccatcac cattaagcgg ccgcactcct
     3121 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     3181 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     3241 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     3301 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     3361 tatttggttt agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct
     3421 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     3481 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     3541 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     3601 cccagtcata gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg
     3661 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3721 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     3841 tccgcgaaat taagcgaaat taactcgaga gatccgctgc attaatgaat cggccaacgc
     3901 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3961 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4021 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4081 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4141 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4201 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4261 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     4321 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4381 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4441 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4501 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4561 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4621 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4681 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4741 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4801 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4861 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4921 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4981 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5041 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5101 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5161 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5221 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5281 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5341 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5401 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5461 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5521 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5581 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5641 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5701 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5761 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5821 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
