LOCUS       sequence-7738-ex        7698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7698)
  AUTHORS   Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K
  TITLE     Vectors for co-expression of an unrestricted number of proteins.
  JOURNAL   Nucleic Acids Res. 2007;35(6):e43. doi: 10.1093/nar/gkm067. Epub 
            2007 Feb 20.
  PUBMED    17311810
REFERENCE   2  (bases 1 to 7698)
  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: "Nucleic 
            Acids Res."; date: "2007"; volume: "35(6):e43. doi"; pages: " 
            10.1093/nar/gkm067. Epub 2007 Feb 20"
COMMENT     Sequence Label: pQLinkGD
FEATURES             Location/Qualifiers
     source          1..7698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(36..50,51..56,57..73,74..80)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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)."
     protein_bind    88..104
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             141..794
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     752..774
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    816..940
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1369..1674)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1471..1490)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2019..2678)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2679..2781)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2959..3083)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3105..3199
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3293..3952
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      4017..4103
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4196..5278)
                     /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     5240..5259
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5279..5356)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5440..5462)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5548..5688
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5603..5622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5703..5720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5874..6462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5954..5973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6633..7424,7425..7493))
                     /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     7256..7275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7494..7598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7666..7684
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
       61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      121 ttaaagagga gaaattaact atgtccccta tactaggtta ttggaaaatt aagggccttg
      181 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
      241 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
      301 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
      361 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
      421 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
      481 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
      541 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
      601 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
      661 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
      721 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
      781 accatcctcc aaaatcggat ctgggtgctg caggtacaag tttgtacaaa aaagctgaac
      841 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
      901 ctacataata ctgtaaaaca caacatatcc agtcactatg aatcaactac ttagatggta
      961 ttagtgacct gtagtcgacc gacagccttc caaatgttct tcgggtgatg ctgccaactt
     1021 agtcgaccga cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc cagcctactc
     1081 gctattgtcc tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga ggtgcgagcc
     1141 tcttttttgt gtgacaaaat aaaaacatct acctattcat atacgctagt gtcatagtcc
     1201 tgaaaatcat ctgcatcaag aacaatttca caactcttat acttttctct tacaagtcgt
     1261 tcggcttcat ctggattttc agcctctata cttactaaac gtgataaagt ttctgtaatt
     1321 tctactgtat cgacctgcag actggctgtg tataagggag cctgacattt atattcccca
     1381 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     1441 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     1501 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc
     1561 actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc
     1621 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc
     1681 agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca
     1741 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca
     1801 tggttgtgct taccagaccg gagatattga catcatatat gccttgagca actgatagct
     1861 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt
     1921 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac
     1981 tgttatctgg cttttagtaa gccggatcca cgcggcgttt acgccccgcc ctgccactca
     2041 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca
     2101 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     2161 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     2221 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     2281 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     2341 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     2401 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     2461 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     2521 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     2581 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     2641 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     2701 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     2761 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
     2821 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
     2881 tattcggcgc aaagtgcgtc gggtgatgct gccaacttag tcgactacag gtcactaata
     2941 ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta ttatgtagtc
     3001 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
     3061 tcagctttct tgtacaaagt ggtaagctta attagctgag cttggactcc tgttgataga
     3121 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     3181 gttttttatt ggtgagaatc caagtaacaa caccatttaa atggagtggt tacaaatgga
     3241 gtggttaatt aaggctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa
     3301 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     3361 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     3421 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     3481 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
     3541 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     3601 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     3661 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     3721 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     3781 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     3841 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     3901 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     3961 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa
     4021 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     4081 acgctctcct gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt
     4141 caaacatgag aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4261 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4321 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4381 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     4441 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     4501 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     4561 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     4621 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     4681 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     4741 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     4801 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     4861 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     4921 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     4981 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5041 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5101 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5161 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5221 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5281 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5341 accattcgat ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat
     5401 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5461 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5521 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     5581 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg
     5641 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     5701 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5761 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5821 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5881 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5941 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6001 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6061 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6301 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7621 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7681 aggccctttc gtcttcac
//
