LOCUS       sequence-7737-ex        5537 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 5537)
  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 5537)
  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: pQLinkG2
FEATURES             Location/Qualifiers
     source          1..5537
                     /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..840
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             840..860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(898..922)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      944..1038
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1132..1791
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1198..1217)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1856..1942
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2035..3117)
                     /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     3079..3098
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3118..3195)
                     /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(3279..3301)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3387..3527
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3442..3461
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3542..3559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3713..4301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3793..3812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4472..5263,5264..5332))
                     /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     5095..5114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5333..5437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5505..5523
                     /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 acatatttga 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 aaagcaggcg
      841 agaatcttta ttttcaggga tccagtcttc gcatgcatga agacgcggcc gcctaggacc
      901 cagctttctt gtacaaagtg gtaagcttaa ttagctgagc ttggactcct gttgatagat
      961 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     1021 ttttttattg gtgagaatcc aagtaacaac accatttaaa tggagtggtt acaaatggag
     1081 tggttaatta aggctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     1141 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1201 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1321 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1621 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1681 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1741 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     1801 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     1861 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1921 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     1981 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2101 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2161 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2221 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2281 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2341 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2401 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2461 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2521 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2581 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2641 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2701 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2761 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2821 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2881 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2941 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3001 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3061 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3121 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     3181 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3241 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3301 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3361 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3421 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc
     3481 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4141 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5461 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5521 ggccctttcg tcttcac
//
