LOCUS       sequence-41850-e        5432 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 5432)
  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 5432)
  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-NEUGRIN
FEATURES             Location/Qualifiers
     source          1..5432
                     /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"
     terminator      889..983
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1027..1686
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1093..1112)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1751..1837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1930..3012)
                     /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     2974..2993
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3013..3090)
                     /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(3174..3196)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3282..3422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3337..3356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3437..3454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3608..4196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4367..5158,5159..5227))
                     /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     4990..5009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5228..5332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5400..5418
                     /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 cagggatccg aggcgcctgg tgccccgccc aggaccctga cgtgggaagc catggagcag
      241 atacggtatt tacatgagga atttccagag tcctggtcag ttcccaggtt ggctgaaggc
      301 tttgatgtca gcactgatgt gatccgaaga gttttaaaaa gcaagttttt acccacattg
      361 gagcagaagc tgaagcagga tcaaaaagtc cttaagaaag ctgggcttgc ccactcgctg
      421 cagcacctcc ggggctctgg aaatacctca aagctgctcc ctgcaggcca ctctgtatca
      481 ggctctttgc ttatgccggg gcatgaagcc tcatctaaag acccaaatca cagcacagct
      541 ttgaaagtga tagagtcaga cactcacagg acaaatacac caaggagaag gaagggaaga
      601 aataaagaaa tccaggacct ggaggagagc tttgtgcctg ttgctgcacc cctaggtcat
      661 ccaagagagc tgcagaagta ctccagtgat tctgagagcc ccagaggaac tggcagtggt
      721 gcgttgccaa gtggtcagaa gctggaggag ttgaaggcag aggagccaga taacttcagc
      781 agcaaagtag tgcagagggg ccgagagttc tttgacagca acgggaactt cctgtacaga
      841 atttgagcgg ccgctcgacc tgcagccaag cttaattagc tgagcttgga ctcctgttga
      901 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
      961 gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa
     1021 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1081 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1141 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1201 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     1261 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1321 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1381 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1441 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1501 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1561 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1621 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1681 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag
     1741 cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1801 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagat tacgtgcagt
     1861 cgatgataag ctgtcaaaca tgagaattgt gcctaatgag tgagctaact tacattaatt
     1921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     2041 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     2101 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     2161 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     2221 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     2281 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     2341 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     2401 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     2461 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     2521 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     2581 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     2641 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     2701 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     2761 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     2821 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     2881 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     2941 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     3001 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     3061 gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca
     3121 cgggtgcgca tgatctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     3181 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3241 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     3301 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     3361 gagtgcacca catgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3421 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4021 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5341 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5401 ataggcgtat cacgaggccc tttcgtcttc ac
//
