LOCUS       sequence-7734-ex        4901 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 4901)
  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 4901)
  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: pQLinkH
FEATURES             Location/Qualifiers
     source          1..4901
                     /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             147..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    180..204
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             204..224
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(262..286)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      308..402
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             496..1155
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(562..581)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1220..1306
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1399..2481)
                     /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     2443..2462
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2482..2559)
                     /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(2643..2665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2751..2891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2806..2825
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2906..2923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3077..3665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3157..3176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3836..4627,4628..4696))
                     /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     4459..4478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4697..4801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4869..4887
                     /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 atgaaacatc accatcacca tcaccatggt gctgcaggta
      181 caagtttgta caaaaaagca ggcgagaatc tttattttca gggatccagt cttcgcatgc
      241 atgaagacgc ggccgcctag gacccagctt tcttgtacaa agtggtaagc ttaattagct
      301 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      361 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagtaa caacaccatt
      421 taaatggagt ggttacaaat ggagtggtta attaaggcta gcttggcgag attttcagga
      481 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      541 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      601 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
      661 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
      721 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
      781 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
      841 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
      901 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
      961 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     1021 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1081 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1141 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     1201 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1261 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cctctagatt
     1321 acgtgcagtc gatgataagc tgtcaaacat gagaattgtg cctaatgagt gagctaactt
     1381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1441 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     1501 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     1561 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     1621 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     1681 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     1741 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     1801 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     1861 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     1921 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     1981 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     2041 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     2101 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     2161 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     2221 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     2281 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     2341 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     2401 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     2461 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     2521 tgccataccg cgaaaggttt tgcaccattc gatggtgtcg gaatttcggg cagcgttggg
     2581 tcctggccac gggtgcgcat gatctagagc tgcctcgcgc gtttcggtga tgacggtgaa
     2641 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2701 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     2761 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     2821 ttgtactgag agtgcaccac atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2881 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3481 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4801 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4861 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
