LOCUS       sequence-41846-e        5588 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 5588)
  AUTHORS   Bussow K, Scheich C, Sievert V, Harttig U, Schultz J, Simon B, Bork 
            P, Lehrach H, Heinemann U
  TITLE     Structural genomics of human proteins--target selection and 
            generation of a public catalogue of expression clones.
  JOURNAL   Microb Cell Fact. 2005 Jul 5;4:21. doi: 10.1186/1475-2859-4-21.
  PUBMED    15998469
REFERENCE   2  (bases 1 to 5588)
  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: "Microb Cell
            Fact."; date: "2005-07-5"; volume: "4:21. doi"; pages: " 
            10.1186/1475-2859-4-21"
COMMENT     Sequence Label: pQTEV-CUTC
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             976..999
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      1045..1139
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1183..1842
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1249..1268)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1907..1993
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2086..3168)
                     /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     3130..3149
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3169..3246)
                     /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(3330..3352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3438..3578
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3493..3512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3593..3610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3764..4352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3844..3863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4523..5314,5315..5383))
                     /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     5146..5165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5384..5488)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5556..5574
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatct aaaaggcagg gggcctcctc tgagcgaaaa
      181 cgagcgcgga taccgtccgg gaaggccgga gcagcaaatg gatttctcat ggaagtttgt
      241 gttgattcag tggaatcagc tgtgaatgca gaaagaggag gtgctgatcg gattgaatta
      301 tgttctggtt tatcagaggg gggaactaca cccagcatgg gtgtccttca agtagtgaag
      361 cagagtgttc agatcccagt ttttgtgatg attcggccac ggggaggtga ttttttgtat
      421 tcagatcgtg aaattgaggt gatgaaggct gacattcgtc ttgccaagct ttatggtgct
      481 gatggtttgg tttttggggc attgactgaa gatggacaca ttgacaaaga gctgtgtatg
      541 tcccttatgg ctatttgccg ccctctgcca gtcactttcc accgagcctt tgacatggtt
      601 catgatccaa tggcagctct ggagaccctc ttaaccttgg gatttgaacg cgtgttgacc
      661 agtggatgtg acagttcagc attagaaggg ctacccctaa taaagcgact cattgagcag
      721 gcaaaaggca ggattgtggt aatgccagga ggtggtataa cagacagaaa tctacaaagg
      781 atccttgagg gttcaggtgc tacagaattc cactgttctg ctcggtctac tagagactcg
      841 ggaatgaagt ttcgaaattc atctgttgcc atgggagcct cactttcttg ctcagaatat
      901 tccctaaagg taacagatgt gaccaaagta aggactttga atgctatcgc aaagaacatc
      961 ctggtggcgg ccgcttggag ccacccgcag ttcgaaaaat aagcggccgc tcgacctgca
     1021 gccaagctta attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac
     1081 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     1141 caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     1201 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1261 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1321 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1381 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1441 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1501 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1561 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1621 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1681 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1741 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     1801 gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     1861 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     1921 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     1981 gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt caaacatgag
     2041 aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     2101 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2161 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     2221 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     2281 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2341 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     2401 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     2461 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     2521 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     2581 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     2641 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     2701 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     2761 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2821 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2881 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2941 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     3001 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     3061 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     3121 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     3181 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     3241 ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat ctagagctgc
     3301 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3361 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3421 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3481 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg cggtgtgaaa
     3541 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     3601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4201 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5521 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5581 gtcttcac
//
