LOCUS       sequence-233154-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Bxb1 integrase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  AUTHORS   Alexander JM, Guan J, Li B, Maliskova L, Song M, Shen Y, Huang B, 
            Lomvardas S, Weiner OD
  TITLE     Live-cell imaging reveals enhancer-dependent Sox2 transcription in 
            the absence of enhancer proximity.
  JOURNAL   Elife. 2019 May 24;8. pii: 41769. doi: 10.7554/eLife.41769.
  PUBMED    31124784
REFERENCE   2  (bases 1 to 6453)
  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: "Elife."; 
            date: "2019-05-24"; volume: "8. pii: 41769. doi"; pages: " 
            10.7554/eLife.41769"
COMMENT     Sequence Label: pCAGGS-Bxb1o_nlsFlag
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..662
                     /label=chicken beta-actin promoter
     intron          663..1672
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1680..1699
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1739..1763
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3318..3341
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(3359..3383)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3475..3494)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3540..3595
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3594..3613)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3956..3972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3956..3972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3969..3991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3980..3996
                     /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(4004..4010,4011..4028,4029..4034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4049..4070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4129..4324
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4175..4310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4330..4464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4367..4386)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4421..4440
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4532..4549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4703..5291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4783..4802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5462..6253,6254..6322))
                     /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     6085..6104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6323..6427)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa
      841 gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct gccgcagggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagan gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1681 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg aggatatcac
     1741 aagtttgtac aaaaaagcag gctttaaagg aaccaattca gtcgactgga tccgccacca
     1801 tgcgagccct cgtggtgatc cggctctcta gagtgaccga tgccactact tcacctgaac
     1861 gacagctgga gtcctgtcag cagctgtgtg ctcagcgcgg ttgggatgtt gtgggggtag
     1921 ctgaagactt ggatgtctcc ggcgccgtag atccttttga tcgcaagcgc aggccaaatt
     1981 tggctcgctg gcttgccttt gaagagcagc ctttcgacgt gatagtggcc tatcgggtgg
     2041 accgactgac aagatctatt cgccatctgc aacagctggt gcattgggca gaagaccaca
     2101 agaaattggt cgtcagcgca accgaagctc attttgatac aaccacccca ttcgccgctg
     2161 tcgtcatcgc cctgatggga acagtcgctc agatggagct ggaggctatc aaagagcgca
     2221 accgctccgc tgcacacttt aatatcaggg caggcaagta caggggctca ctcccgccgt
     2281 ggggttatct tcccacaagg gtggatggcg agtggagact cgtgccggac cctgtgcaaa
     2341 gggagcgcat ccttgaagtt taccataggg ttgtagataa ccacgagcct ctccacctcg
     2401 tggcccatga ccttaataga cggggagtgt tgagtcccaa ggactacttc gcccagctgc
     2461 aggggcggga gccccagggt agggagtggt ctgcaactgc cctgaagcga tccatgatca
     2521 gcgaagccat gcttggatat gccaccctca acggcaagac agtcagggac gatgacggcg
     2581 caccactggt tagggctgaa cccattctga ccagggaaca gctggaagct ttgcgcgcag
     2641 agttggtgaa gacctcccgg gcaaagcccg ccgtctctac tccgtccctg ctcctgagag
     2701 tattgttctg tgcagtgtgc ggcgagcctg cctataagtt tgcaggaggt ggccgcaaac
     2761 acccacgcta ccgctgccgc tccatgggct ttccaaagca ctgtggtaat ggcacagttg
     2821 ccatggccga gtgggacgct ttctgtgagg agcaggtgct tgacctcctg ggtgatgctg
     2881 agcgcttgga gaaggtgtgg gttgccggtt ccgatagcgc agtagagttg gccgaggtta
     2941 acgctgagct tgtggatctg actagcctga tcgggtctcc agcctacagg gccggctctc
     3001 ctcaaagaga ggcactcgac gcacggatcg ccgctctggc cgcccgacag gaagagctgg
     3061 aggggctgga agctcgccca agcggatggg aatggaggga gacaggccag aggtttgggg
     3121 attggtggcg cgaacaggac accgcagcca aaaacacctg gctccgatct atgaacgtta
     3181 gactcacttt tgatgtcagg ggaggtctga ctcggacaat agacttcggc gacctgcagg
     3241 agtatgaaca gcacctcaga ttgggcagtg tggtcgaacg attgcacaca ggtatgtcta
     3301 agaaaaagcg gaaagtggac tacaaagacg atgacgacaa atgactcgag atatctagac
     3361 ccagctttct tgtacaaagt ggtgatatcg cggccgcact cctcaggtgc aggctgccta
     3421 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3481 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3541 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3601 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3661 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3721 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3781 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3841 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     3901 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     3961 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4021 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4081 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     4141 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4201 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4261 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4321 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4381 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4441 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4501 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4561 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4621 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5401 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5461 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5521 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5581 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5641 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5701 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5761 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5821 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5881 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5941 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6001 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6061 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6421 ttccgcgcac atttccccga aaagtgccac ctg
//
