LOCUS       sequence-41903-e        5348 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 5348)
  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 5348)
  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-EPSTI1
FEATURES             Location/Qualifiers
     source          1..5348
                     /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             121..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             602..619
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     terminator      805..899
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             943..1602
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1009..1028)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1667..1753
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1846..2928)
                     /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     2890..2909
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2929..3006)
                     /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(3090..3112)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3198..3338
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3253..3272
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3353..3370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3524..4112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3604..3623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4283..5074,5075..5143))
                     /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     4906..4925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5144..5248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5316..5334
                     /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 aactatgaaa
      121 catcaccatc accatcacca tagcgattac gacatcccca ctactgagaa tctttatttt
      181 cagggatccc aatctaaata caagcaaaag ctaaaaagag aagaatctgt aagaatcaag
      241 aaggaagctg aagaagctga actccaaaaa atgaaggcaa ttcagagaga gaagagcaat
      301 aaactggagg agaaaaaaag acttcaagaa aaccttagaa gagaagcatt tagagagcat
      361 cagcaataca aaaccgctga gttcttgagc aaactgaaca cagagtcgcc agacagaagt
      421 gcctgtcaaa gtgctgtttg tggcccacaa tcctcaacat ggaaacttcc tatcctgcct
      481 agggatcaca gctgggccag aagctgggct tacagagatt ctctaaaggc agaagaaaac
      541 agaaaattgc aaaagatgaa ggatgaacaa catcaaaaga gtgaattact ggaactgaaa
      601 cggcagcagc aagagcaaga aagagccaaa atccaccaga ctgaacacag gagggtaaat
      661 aatgcttttc tggaccgact ccaaggcaaa agtcaaccag gtggcctcga gcaatctgga
      721 ggctgttgga atatgaatag cggtaacagc tggggtatat gagcggccgc tcgacctgca
      781 gccaagctta attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac
      841 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
      901 caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
      961 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1021 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1081 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1141 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1201 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1261 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1321 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1381 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1441 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1501 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     1561 gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     1621 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     1681 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     1741 gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt caaacatgag
     1801 aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     1861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1921 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1981 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     2041 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2101 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     2161 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     2221 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     2281 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     2341 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     2401 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     2461 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     2521 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2581 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2641 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2701 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2761 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     2821 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     2881 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2941 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     3001 ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat ctagagctgc
     3061 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3121 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3181 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3241 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg cggtgtgaaa
     3301 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     3361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3961 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5281 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5341 gtcttcac
//
