LOCUS       sequence-7736-ex        4823 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 4823)
  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 4823)
  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: pQLinkN
FEATURES             Location/Qualifiers
     source          1..4823
                     /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)"
     protein_bind    complement(184..208)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      230..324
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             418..1077
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(484..503)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1142..1228
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1321..2403)
                     /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     2365..2384
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2404..2481)
                     /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(2565..2587)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2673..2813
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2728..2747
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2828..2845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2999..3587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3079..3098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3758..4549,4550..4618))
                     /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     4381..4400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4619..4723)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4791..4809
                     /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 atgggatcca gtcttcgcat gcatgaagac gcggccgcct
      181 aggacccagc tttcttgtac aaagtggtaa gcttaattag ctgagcttgg actcctgttg
      241 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
      301 cgggcgtttt ttattggtga gaatccaagt aacaacacca tttaaatgga gtggttacaa
      361 atggagtggt taattaaggc tagcttggcg agattttcag gagctaagga agctaaaatg
      421 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
      481 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
      541 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
      601 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
      661 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
      721 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
      781 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
      841 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
      901 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
      961 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1021 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     1081 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     1141 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1201 ggtgaacgct ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa
     1261 gctgtcaaac atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     1321 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1381 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     1441 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     1501 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     1561 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     1621 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     1681 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     1741 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     1801 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     1861 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     1921 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     1981 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     2041 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     2101 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     2161 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     2221 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     2281 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     2341 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     2401 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     2461 tttgcaccat tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc
     2521 atgatctaga gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2581 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2641 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2701 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2761 acatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3421 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4741 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4801 tcacgaggcc ctttcgtctt cac
//
