LOCUS       sequence-398140-        5199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose-induced expression of His-tagged biliverdin-binding serpin
            of Boana punctata treefrog (BpBBS) in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5199)
  AUTHORS   Manoilov KY, Ghosh A, Almo SC, Verkhusha VV
  TITLE     Structural and Functional Characterization of a Biliverdin-Binding 
            Near-Infrared Fluorescent Protein From the Serpin Superfamily.
  JOURNAL   J Mol Biol. 2022 Jan 30;434(2):167359. doi: 
            10.1016/j.jmb.2021.167359. Epub 2021 Nov 16.
  PUBMED    34798132
REFERENCE   2  (bases 1 to 5199)
  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: "J Mol 
            Biol."; date: "2022-01-30"; volume: "434(2):167359. doi"; pages: " 
            10.1016/j.jmb.2021.167359. Epub 2021 Nov 16"
COMMENT     Sequence Label: pBAD-BpBBS
FEATURES             Location/Qualifiers
     source          1..5199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             236..253
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1508..1525)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1678..1764
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1856..1883
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1902..1993
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1994..2062,2063..2854)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2212..2231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3025..3613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3514..3533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3767..3784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3799..3939
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3865..3884)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4025..4047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4203..5081)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5108..5199,1..193)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
ORIGIN
        1 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
       61 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
      121 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct
      181 actgtttctc catacccgtt ttttgggcta acaggaggaa ttaaccatgg ggagccacca
      241 tcaccatcac catggcagat ctcaccacga ggagggacac cacgaccacg aggatctgaa
      301 ggacgatcac gacccattcc tgcccgagga tcacaagaag gccctgttcg tgtaccagaa
      361 gcccgccctg aacaatatca acttcgcctt taagatgtat agacagctgg ccagggacca
      421 ccctacagag aatatcgtga tctccccagt gtctatcagc tccgccctgg ccctgctgag
      481 cctgggagca aagggacaca cccactccca gatcgtggag agactgggct acaacacatc
      541 tgagatccct gagcagcaga tccacgagag cttccacaag cagctggacg tggtggacga
      601 taaggacagg gatctggagt tcgagcacgg caatgccctg tttacctgca aggagcacaa
      661 gatccaccag acattcctgg acgatgccaa gaagttttac cactctgaag tgatcccaac
      721 cgacttcaag aacacagagg aggccaagaa ccagatcaat agctatgtgg agaagtccac
      781 ccacggcaag atcacaaata tcctggatag cgtggaccag gatgccatga tcgccctgat
      841 caactttatc tatctgcgcg ccaattggca gcaccctttc gacgagaagc tgaccaagga
      901 gggcgatttt cacgtggaca aggataccac agtgaaggtg ccattcatgc ggagaagggg
      961 catctacaag atggcctata ccgacgatat catcatggtg acaatcccct acaacggctc
     1021 cgtggagatg tttctggcca tgacaaagat gggcaagctg tccgagctgg agcagaacct
     1081 gaatcgcgag cggtctctga agtggcggga gatcatgcag tatcagctga tcgacctgag
     1141 cctgcccaag ctgtctgtga gcggcatcct gaacctgaag gagaccctgt ccaagctggg
     1201 catcgtggac gtgttcagca atcacgccga cctgtctggc atcacagatg agtctcacct
     1261 gaaggtgagc aaggccatcc acaaggccat gatgtctttt gacgagcacg gaaccgaggc
     1321 agcaccagca acagcagcag aggcagatcc tctgatgctg ccacctcact tcaagtttga
     1381 ctaccctttc atctttcgcg tgcaggatct gaaaaccaag aacccactgc tggtgggccg
     1441 gatcgccaat ccccagaagt gagaattcga agcttggctg ttttggtgga tgagagaaga
     1501 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1561 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1621 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     1681 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1741 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     1801 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     1861 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     1921 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1981 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2041 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2101 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2161 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2221 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2281 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2341 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2401 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2461 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2521 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2581 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2641 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2701 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2761 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2821 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2881 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2941 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3001 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3061 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3721 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3781 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3841 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3901 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3961 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4021 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4081 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca aatggacgaa
     4141 gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc
     4201 aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     4261 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     4321 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     4381 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     4441 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     4501 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     4561 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     4621 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     4681 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     4741 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     4801 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     4861 aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     4921 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     4981 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     5041 gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca
     5101 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
     5161 tggctcttct cgctaaccaa accggtaacc ccgcttatt
//
