LOCUS       sequence-14467-e        5714 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 5714)
  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 5714)
  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-PRKRA
FEATURES             Location/Qualifiers
     source          1..5714
                     /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      1171..1265
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1309..1968
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1375..1394)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2033..2119
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2212..3294)
                     /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     3256..3275
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3295..3372)
                     /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(3456..3478)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3564..3704
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3619..3638
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3719..3736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3890..4478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3970..3989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4649..5440,5441..5509))
                     /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     5272..5291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5510..5614)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5682..5700
                     /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 cagggatcct cccagagcag gcaccgcgcc gaggccccgc cgctggagcg cgaggacagt
      241 gggaccttca gtttggggaa gatgataaca gctaagccag ggaaaacacc gattcaggta
      301 ttacacgaat acggcatgaa gaccaagaac atcccagttt atgaatgtga aagatctgat
      361 gtgcaaatac acgtgcccac tttcaccttc agagtaaccg ttggtgacat aacctgcaca
      421 ggtgaaggta caagtaagaa gctggcgaaa catagagctg cagaggctgc cataaacatt
      481 ttgaaagcca atgcaagtat ttgctttgca gttcctgacc ccttaatgcc tgacccttcc
      541 aagcaaccaa agaaccagct taatcctatt ggttcattac aggaattggc tattcatcat
      601 ggctggagac ttcctgaata taccctttcc caggagggag gacctgctca taagagagaa
      661 tatactacaa tttgcaggct agagtcattt atggaaactg gaaagggggc atcaaaaaag
      721 caagccaaaa ggaatgctgc tgagaaattt cttgccaaat ttagtaatat ttctccagag
      781 aaccacattt ctttaacaaa tgtagtagga cattctttag gatgtacttg gcattccttg
      841 aggaattctc ctggtgaaaa gatcaactta ctgaaaagaa gcctcctcag tattccaaat
      901 acagattaca tccagctgct tagtgaaatt gccaaggaac aaggttttaa tataacatat
      961 ttggatatag atgaactgag cgccaatgga caatatcaat gtcttgctga actgtccacc
     1021 agccccatca cagtctgtca tggctccggt atctcctgtg gcaatgcaca aagtgatgca
     1081 gctcacaatg ctttgcagta tttaaagata atagcagaaa gaaagtgagc ggccgctcga
     1141 cctgcagcca agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct
     1201 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     1261 agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1321 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1381 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1441 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1501 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1561 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1621 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1681 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1741 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1801 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1861 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1921 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
     1981 agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa
     2041 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2101 tctcctgagt aggacaaatc cgccctctag attacgtgca gtcgatgata agctgtcaaa
     2161 catgagaatt gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     2221 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2281 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2341 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2401 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     2461 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     2521 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     2581 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     2641 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     2701 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     2761 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2821 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2881 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2941 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     3001 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     3061 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     3121 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     3181 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3241 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3301 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca
     3361 ttcgatggtg tcggaatttc gggcagcgtt gggtcctggc cacgggtgcg catgatctag
     3421 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3481 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3541 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     3601 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cacatgcggt
     3661 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     3721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4081 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4321 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4561 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4621 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4681 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4741 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4801 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4861 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4921 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4981 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5041 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5101 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5161 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5221 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5281 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5341 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5401 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5461 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5641 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5701 cctttcgtct tcac
//
