LOCUS       sequence-41883-e        5951 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 5951)
  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 5951)
  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-ILKAP
FEATURES             Location/Qualifiers
     source          1..5951
                     /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      1408..1502
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1546..2205
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1612..1631)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2270..2356
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2449..3531)
                     /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     3493..3512
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3532..3609)
                     /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(3693..3715)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3801..3941
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3856..3875
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3956..3973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4886..5677,5678..5746))
                     /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"
     promoter        complement(5747..5851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5919..5937
                     /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 acctcttcgg ggacctgccg gagcccgagc gctcgccgcg cccggctgcc
      241 gggaaagaag ctcagaaagg acccctgctc tttgatgacc tccctccggc cagcagtact
      301 gactcaggat cagggggacc tttgcttttt gatgatctcc cacccgctag cagtggcgat
      361 tcaggttctc ttgccacatc aatatcccag atggtaaaga ctgaagggaa aggagcaaag
      421 agaaaaacct ccgaggaaga gaagaatggc agtgaagagc ttgtggaaaa gaaagtttgt
      481 aaagcctctt cggtgatctt tggtctgaag ggctatgtgg ctgagcggaa gggtgagagg
      541 gaggagatgc aggatgccca cgtcatcctg aacgacatca ccgaggagtg taggccccca
      601 tcgtccctca ttactcgggt ttcatatttt gctgtttttg atggacatgg aggaattcga
      661 gcctcaaaat ttgctgcaca gaatttgcat caaaacttaa tcagaaaatt tcctaaagga
      721 gatgtaatca gtgtagagaa aaccgtgaag agatgccttt tggacacttt caagcatact
      781 gatgaagagt tccttaaaca agcttccagc cagaagcctg cctggaaaga tgggtccact
      841 gccacgtgtg ttctggctgt agacaacatt ctttatattg ccaacctcgg agatagtcgg
      901 gcaatcttgt gtcgttataa tgaggagagt caaaaacatg cagccttaag cctcagcaaa
      961 gagcataatc caactcagta tgaagagcgg atgaggatac agaaggctgg aggaaacgtc
     1021 agggatgggc gtgttttggg cgtgctagag gtgtcacgct ccattgggga cgggcagtac
     1081 aagcgctgcg gtgtcacctc tgtgcccgac atcagacgct gccagctgac ccccaatgac
     1141 aggttcattt tgttggcctg tgatgggctc ttcaaggtct ttaccccaga agaagccgtg
     1201 aacttcatct tgtcctgtct cgaggatgaa aagatccaga cccgggaagg gaagtccgca
     1261 gccgacgccc gctacgaagc agcctgcaac aggctggcca acaaggcggt gcagcggggc
     1321 tcggccgaca acgtcactgt gatggtggtg cggatagggc actgagcggc cgctcgacct
     1381 gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
     1441 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     1501 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1561 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1621 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1681 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1741 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1801 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1861 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1921 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1981 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2041 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2101 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2161 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     2221 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     2281 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2341 cctgagtagg acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat
     2401 gagaattgtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2461 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2521 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2581 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2641 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     2701 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     2761 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     2821 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     2881 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     2941 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     3001 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     3061 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     3121 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     3181 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     3241 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     3301 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     3361 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     3421 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     3481 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3541 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc
     3601 gatggtgtcg gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc
     3661 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3721 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3781 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3841 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccac atgcggtgtg
     3901 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3961 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4021 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4561 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4741 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4801 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4861 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4921 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4981 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5041 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5101 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5161 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5221 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5281 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5341 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5401 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5461 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5521 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5581 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5641 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5701 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5761 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5821 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5881 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5941 ttcgtcttca c
//
