LOCUS       sequence-388613-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Bxb1 integrase fused with CFP for fluorescence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4526)
  AUTHORS   Pandey A, Rodriguez ML, Poole W, Murray RM
  TITLE     Characterization of Integrase and Excisionase Activity in a 
            Cell-Free Protein Expression System Using a Modeling and Analysis 
            Pipeline.
  JOURNAL   ACS Synth Biol. 2023 Feb 17;12(2):511-523. doi: 
            10.1021/acssynbio.2c00534. Epub 2023 Jan 30.
  PUBMED    36715625
REFERENCE   2  (bases 1 to 4526)
  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"; doi: 
            "10.1021/acssynbio.2c00534"; journalName: "ACS Synth Biol"; date: 
            "2023-02-17- 17"; volume: "12"; issue: "2"; pages: "511-523"
COMMENT     Sequence Label: pTet-Bxb1-CFP
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..57
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    64..82
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             163..171
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             178..1677
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISGAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAVRQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     CDS             1717..1734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1762..1783)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2023..2042)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2370..2391
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2447..2467
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2468..2525
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2549..2566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2776..3363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2856..2875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3451..3545)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3566..4225)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTIQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4140..4159
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4226..4328)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4505..4526,1..23))
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 tagctttcgc taaggatgat ttctggaatt ttctagagtc cctatcagtg atagagattg
       61 acatccctat cagtgataga gatactgagc acgggcccaa gttcacttaa aaaggagatc
      121 aacaatgaaa gcaattttcg tactgaaaca tcttaatcat gctaaggagg ttttctaatg
      181 agagccctgg tagtcatccg cctgtcccgc gtcaccgatg ctacgacttc accggagcgt
      241 cagctggagt cttgccagca gctctgcgcc cagcgcggct gggacgtcgt cggggtagcg
      301 gaggatctgg acgtctccgg ggcggtcgat ccgttcgacc ggaagcgcag accgaacctg
      361 gcccggtggc tagcgttcga ggagcaaccg tttgacgtga tcgtggcgta ccgggtagat
      421 cggttgaccc gatcgatccg gcatcttcag cagctggtcc actgggccga ggaccacaag
      481 aagctggtcg tctccgcgac cgaagcgcac ttcgatacga cgacgccgtt tgcggcggtc
      541 gtcatcgcgc ttatgggaac ggtggcgcag atggaattag aagcgatcaa agagcggaac
      601 cgttcggctg cgcatttcaa tatccgcgcc gggaaatacc gaggatccct gccgccgtgg
      661 ggatacctgc ctacgcgcgt ggacggggag tggcggctgg tgccggaccc tgtgcagcga
      721 gagcgcatcc tcgaggtgta tcaccgcgtc gtcgacaacc acgagccgct gcatctggtg
      781 gcccacgacc tgaaccggcg tggtgtcctg tcgccgaagg actacttcgc gcagctgcaa
      841 ggccgcgagc cgcagggccg ggagtggtcg gctaccgcgc tgaagcgatc gatgatctcc
      901 ggggcgatgc tcgggtacgc gactctgaac ggtaagaccg tccgagacga cgacggagcc
      961 ccgctggtgc gggctgagcc gatcctgacc cgtgagcagc tggaggcgct gcgcgccgag
     1021 ctcgtgaaga cctcccgggc gaagcccgcg gtgtctaccc cgtcgctgct gctgcgggtg
     1081 ttgttctgcg cggtgtgcgg ggagcccgcg tacaagttcg ccgggggagg acgtaagcac
     1141 ccgcgctacc gctgccgctc gatggggttc ccgaagcact gcgggaacgg cacggtggcg
     1201 atggccgagt gggacgcgtt ctgcgaggag caggtactgg atctgctcgg ggacgcggag
     1261 cgtctggaga aagtctgggt agcgggctcg gactccgcgg tcgaactcgc ggaggtgaac
     1321 gcggagctgg tggacctgac gtcgctgatc ggctccccgg cctaccgggc gggctctccg
     1381 cagcgagaag cactggatgc ccgtattgcg gcgctggccg tgcggcaaga ggagctggag
     1441 ggcctggagg ctcgcccgtc tggctgggag tggcgcgaga ccgggcagcg gttcggggac
     1501 tggtggcggg agcaggacac cgcggcaaag aacacctggc ttcggtcgat gaacgttcgg
     1561 ctgacgttcg acgtccgcgg cgggctgact cgcacgatcg acttcgggga tcttcaggag
     1621 tacgagcagc atctcaggct cggcagcgtg gtcgaacggc tacacaccgg gatgtcggga
     1681 agcggaagtg gatccggtag tggcagcggc agcatgcatc atcaccacca ccatgagctt
     1741 ttcactggcg ttgttcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1801 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1861 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc
     1921 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1981 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2041 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2101 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc
     2161 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     2221 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2281 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2341 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2401 gggatctaac tcgagcaaag cccgccgaaa ggcgggcttt tctgtatact agtagcggcc
     2461 gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
     2521 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat cctagctagg gcgttcggct
     2641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2941 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3001 gtaggtcgtt cgctccaagc tgggctgtat gcacgaaccc cccgttcagc ccgaccgctg
     3061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3181 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3301 cgctggtagc ggtggttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3361 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3421 taagggattt tggtcatgac ttgtgcttgg attctcacca ataaaaaacg cccggcggca
     3481 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     3541 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     3601 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     3661 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     3721 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     3781 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     3841 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3901 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3961 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     4021 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     4081 ttacggtctt taaaaaggcc gtaatatcca gctgaatggt ctggttatag gtacattgag
     4141 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4201 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     4261 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4321 gccgatcaac gtctcatttt cgccagatat cgacgtctaa gaaaccatta ttatcatgac
     4381 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg agtgccacct
     4441 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4501 cagaatttca gataaaaaaa aatcct
//
