LOCUS       sequence-41848-e        5438 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 5438)
  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 5438)
  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-VPS28
FEATURES             Location/Qualifiers
     source          1..5438
                     /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             complement(220..237)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      895..989
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1033..1692
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1099..1118)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1757..1843
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1936..3018)
                     /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     2980..2999
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3019..3096)
                     /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(3180..3202)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3288..3428
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3343..3362
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3443..3460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3614..4202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3694..3713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4373..5164,5165..5233))
                     /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     4996..5015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5234..5338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5406..5424
                     /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 cagggatctt ttcatgggat cccagccacg ccgggcatag gagcccctgg gaacaagccg
      241 gagctgtatg aggaagtgaa gttgtacaag aacgcccggg agagggagaa gtacgacaac
      301 atggcagagc tgtttgcggt ggtgaagaca atgcaagccc tggagaaggc ctacatcaag
      361 gactgtgtct cccccagcga gtacactgca gcctgctccc ggctcctggt ccaatacaaa
      421 gctgccttca ggcaggtcca gggctcagaa atcagctcta ttgacgaatt ctgccgcaag
      481 ttccgcctgg actgcccgct ggccatggag cggatcaagg aggaccggcc catcaccatc
      541 aaggacgaca agggcaacct caaccgctgc atcgcagacg tggtctcgct cttcatcacg
      601 gtcatggaca agctgcgcct ggagatccgc gccatggatg agatccagcc cgacctgcga
      661 gagctgatgg agaccatgca ccgcatgagc cacctcccac ccgactttga gggccgccag
      721 acggtcagcc agtggctgca gaccctgagc ggcatgtcgg cgtcagatga gctggacgac
      781 tcacaggtgc gtcagatgct gttcgacctg gagtcagcct acaacgcctt caaccgcttc
      841 ctgcatgcct gagcggccgc tcgacctgca gccaagctta attagctgag cttggactcc
      901 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
      961 gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt ttcaggagct
     1021 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1081 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1141 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1201 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
     1261 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     1321 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1381 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     1441 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
     1501 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     1561 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     1621 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     1681 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg ggtaatgact
     1741 ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1801 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct ctagattacg
     1861 tgcagtcgat gataagctgt caaacatgag aattgtgcct aatgagtgag ctaacttaca
     1921 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     2041 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     2101 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     2161 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     2221 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     2281 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     2341 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     2401 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     2461 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     2521 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     2581 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     2641 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     2701 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     2761 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     2821 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     2881 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     2941 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     3001 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     3061 cataccgcga aaggttttgc accattcgat ggtgtcggaa tttcgggcag cgttgggtcc
     3121 tggccacggg tgcgcatgat ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     3181 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3241 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     3301 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     3361 tactgagagt gcaccacatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3421 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4021 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5341 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5401 ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
