LOCUS       sequence-242357-        4256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible vector encoding the N-terminal gp41-1 split fragment 
            with N-terminally His-tagged GB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4256)
  AUTHORS   Beyer HM, Iwai H
  TITLE     Off-pathway-sensitive protein splicing screening based on a 
            toxin/antitoxin system.
  JOURNAL   Chembiochem. 2019 Apr 8. doi: 10.1002/cbic.201900139.
  PUBMED    30963690
REFERENCE   2  (bases 1 to 4256)
  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: 
            "Chembiochem. 2019 Apr 8. doi: 10.1002/cbic.201900139."
COMMENT     Sequence Label: pBHDuet40
FEATURES             Location/Qualifiers
     source          1..4256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..48
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    49..73
                     /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)."
     CDS             129..146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             156..173
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        687..705
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    706..730
                     /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)."
     CDS             839..883
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(921..939)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      935..982
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1212..2027)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1212..2027)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1935..1954
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2028..2119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2135..2884)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2145..2162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2385..2404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3080..4162)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4124..4143
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4163..4240)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tcccttatgc gactcctgca ttaggaaatt aatacgactc actatagggg aattgtgagc
       61 ggataacaat tcccctgtag aaataatttt gtttaacttt aataaggaga tataccatgg
      121 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgc
      181 agtacaaact tatcctgaac ggtaaaaccc tgaaaggtga aaccaccacc gaagctgttg
      241 acgctgctac cgcggaaaaa gttttcaaac agtacgctaa cgacaacggt gttgacggtg
      301 aatggaccta cgacgacgct accaaaacct acaccgtaac ggaaggatcc tgcctggatc
      361 tgaaaacgca ggtgcagacc ccacagggca tgaaagagat atccaacatt caggtgggcg
      421 atcttgttct gagcaacacc ggctataacg aggtgctgaa cgtgtttccg aaaagcaaaa
      481 agaaaagcta taaaattacc ctggaagatg gcaaagaaat tatttgtagc gaagaacatc
      541 tgttcccgac ccagaccggc gaaatgaaca tcagcggcgg tctgaaagag ggcatgtgcc
      601 tgtatgtgaa agaataagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      661 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      721 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatggcaga
      781 tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa
      841 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
      901 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      961 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
     1021 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     1081 ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact tttcggggaa
     1141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1201 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1261 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1321 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1381 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1441 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1501 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1561 attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa
     1621 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1681 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     1741 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1801 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1861 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     1921 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     1981 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
     2041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2101 atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct cactgactcg
     2161 ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg
     2221 ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag caaagcaccg
     2281 gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2341 aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2401 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2461 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gttggtatct
     2521 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2581 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2641 atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg aagttatgca
     2701 cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca acccacttac
     2761 cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg
     2821 tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga acagctattc
     2881 cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2941 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     3001 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     3061 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3121 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     3181 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     3241 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     3301 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     3361 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     3421 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     3481 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3541 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3601 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3661 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     3721 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     3781 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     3841 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     3901 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     3961 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     4021 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     4081 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     4141 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     4201 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctc
//
