LOCUS       sequence-14359-e        6254 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 6254)
  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 6254)
  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-PEPD
FEATURES             Location/Qualifiers
     source          1..6254
                     /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      1711..1805
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1849..2508
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1915..1934)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2573..2659
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2752..3834)
                     /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     3796..3815
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3835..3912)
                     /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(3996..4018)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4104..4244
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4159..4178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4259..4276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4430..5018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4510..4529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5189..5980,5981..6049))
                     /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     5812..5831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6050..6154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6222..6240
                     /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 cggcggccac cggaccctcg ttttggctgg ggaatgaaac cctgaaggtg
      241 ccgctggcgc tctttgcctt gaaccggcag cgcctgtgtg agcggctgcg gaagaaccct
      301 gctgtgcagg ccggctccat cgtggtcctg cagggcgggg aggagactca gcgctactgc
      361 accgacaccg gggtcctctt ccgccaggag tccttctttc actgggcgtt cggtgtcact
      421 gagccaggct gctatggtgt catcgatgtt gacactggga agtcgaccct gtttgtgccc
      481 aggcttcctg ccagccatgc cacctggatg ggaaagatcc attccaagga gcacttcaag
      541 gagaagtatg ccgtggacga cgtccagtac gtagatgaga ttgccagcgt cctgacgtca
      601 cagaagccct ctgtcctcct cactttgcgt ggcgtcaaca cggacagcgg cagtgtctgc
      661 agggaggcct cctttgacgg catcagcaag ttcgaagtca acaataccat tcttcaccca
      721 gagatcgttg agtgccgagt gtttaagacg gatatggagc tggaggttct gcgctatacc
      781 aataaaatct ccagcgaggc ccaccgtgag gtaatgaagg ctgtaaaagt gggaatgaaa
      841 gaatatgagt tggaaagcct cttcgagcac tactgctact cccggggcgg catgcgccac
      901 agctcctaca cctgcatctg cggcagtggt gagaactcag ccgtgctaca ctacggacac
      961 gccggagctc ccaacgaccg aacgatccag aatggggata tgtgcctgtt cgacatgggc
     1021 ggtgagtatt actgcttcgc ttccgacatc acctgctcct ttcccgccaa cggcaagttc
     1081 actgcagacc agaaggccgt ctatgaggca gtgctgcgga gctcccgtgc cgtcatgggt
     1141 gccatgaagc caggtgtctg gtggcctgac atgcaccgcc tggctgaccg catccacctg
     1201 gaggagctgg cccacatggg catcctgagc ggcagcgtgg acgccatggt ccaggctcac
     1261 ctgggggccg tgtttatgcc tcacgggctt ggccacttcc tgggcattga cgtgcacgac
     1321 gtgggaggct acccagaggg cgtggagcgc atcgacgagc ccggcctgcg gagcctgcgc
     1381 actgcacggc acctgcagcc aggcatggtg ctcaccgtgg agccgggcat ctacttcatc
     1441 gaccacctcc tggatgaggc cctggcggac ccggcccgcg cctccttctt taaccgcgag
     1501 gtcctgcagc gctttcgcgg ttttggcggg gtccgcatcg aggaggacgt cgtggtgact
     1561 gacagcggca tagagctgct gacctgcgtg ccccgcactg tggaagagat tgaagcatgc
     1621 atggcaggct gtgacaaggc ctttaccccc ttctctggcc ccaagtaggc ggccgctcga
     1681 cctgcagcca agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct
     1741 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     1801 agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1861 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1921 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1981 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     2041 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     2101 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     2161 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     2221 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     2281 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     2341 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     2401 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     2461 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
     2521 agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa
     2581 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2641 tctcctgagt aggacaaatc cgccctctag attacgtgca gtcgatgata agctgtcaaa
     2701 catgagaatt gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     2761 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2821 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2881 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2941 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     3001 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     3061 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     3121 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     3181 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     3241 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     3301 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     3361 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     3421 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     3481 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     3541 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     3601 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     3661 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     3721 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3781 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3841 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca
     3901 ttcgatggtg tcggaatttc gggcagcgtt gggtcctggc cacgggtgcg catgatctag
     3961 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4021 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4081 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     4141 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cacatgcggt
     4201 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     4261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4861 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6181 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6241 cctttcgtct tcac
//
