LOCUS       sequence-14483-e        5822 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 5822)
  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 5822)
  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-PTK9L
FEATURES             Location/Qualifiers
     source          1..5822
                     /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      1279..1373
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1417..2076
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1483..1502)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2141..2227
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2320..3402)
                     /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     3364..3383
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3403..3480)
                     /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(3564..3586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3672..3812
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3727..3746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3827..3844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4757..5548,5549..5617))
                     /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     5380..5399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5618..5722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5790..5808
                     /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 cgcaccaaac gggcatccac gccacggaag agctgaagga attctttgcc
      241 aaggcacggg ctggctctgt gcggctcatc aaggttgtga ttgaggacga gcagctcgtg
      301 ctgggtgcct cgcaggagcc agtaggccgc tgggatcagg actatgacag ggccgtgctg
      361 ccactgctgg acgcccagca gccctgctac ctgctctacc gcctcgactc acagaatgct
      421 cagggcttcg aatggctctt cctcgcctgg tcgcctgata actcccccgt gcggctgaag
      481 atgctgtacg cggccacgcg ggccacagtg aaaaaggagt ttggaggtgg ccacatcaag
      541 gatgagctct tcgggactgt gaaggatgac ctctcttttg ctgggtacca gaaacacctg
      601 tcgtcctgtg cggcacctgc cccgctgacc tcggctgaga gagagctcca gcagatccgc
      661 attaacgagg tgaagacaga gatcagtgtg gaaagcaagc accagaccct gcagggcctc
      721 gccttccccc tgcagcctga ggcccagcgg gcactccagc agctcaagca gaaaatggtc
      781 aactacatcc agatgaagct ggacctagag cgggaaacca ttgagctggt gcacacagag
      841 cccacggatg tggcccagct gccctcccgg gtgccccgag atgctgcccg ctaccacttc
      901 ttcctctaca agcacaccca tgagggcgac ccccttgagt ctgtagtgtt catctactcc
      961 atgccggggt acaagtgcag catcaaggag cgaatgctct actccagctg caagagccgc
     1021 ctcctcgact ccgtggagca ggacttccat ctggagatcg ccaagaaaat tgagattggc
     1081 gatggggcag agctgacggc agagttcctc tacgacgagg tgcaccccaa gcaacacgcc
     1141 ttcaagcagg ccttcgccaa gcccaagggc ccagggggca agcggggcca taagcgcctc
     1201 atccgcggcc cgggtgaaaa tggggatgac agctgagcgg ccgctcgacc tgcagccaag
     1261 cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1321 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct
     1381 agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1441 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1501 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1561 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     1621 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     1681 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1741 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1801 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1861 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1921 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1981 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2041 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     2101 ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac gaaaggctca
     2161 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     2221 gacaaatccg ccctctagat tacgtgcagt cgatgataag ctgtcaaaca tgagaattgt
     2281 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2341 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2401 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     2461 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     2521 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     2581 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     2641 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     2701 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     2761 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     2821 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     2881 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     2941 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3001 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     3061 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     3121 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     3181 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     3241 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     3301 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     3361 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     3421 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc
     3481 ggaatttcgg gcagcgttgg gtcctggcca cgggtgcgca tgatctagag ctgcctcgcg
     3541 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3601 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3661 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     3721 actatgcggc atcagagcag attgtactga gagtgcacca catgcggtgt gaaataccgc
     3781 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5041 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5341 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5761 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     5821 ac
//
