LOCUS       sequence-14482-e        5318 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 5318)
  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 5318)
  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-PDAP1
FEATURES             Location/Qualifiers
     source          1..5318
                     /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      775..869
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             913..1572
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(979..998)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1637..1723
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1816..2898)
                     /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     2860..2879
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2899..2976)
                     /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(3060..3082)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3168..3308
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3223..3242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3323..3340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3494..4082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3574..3593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4253..5044,5045..5113))
                     /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     4876..4895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5114..5218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5286..5304
                     /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 cagggatccc ctaaaggagg aagaaaggga ggccacaaag gccgggcgag gcagtataca
      241 agccctgagg agatcgacgc gcagctgcag gctgagaagc agaaggccag ggaagaagag
      301 gagcaaaaag aaggtggaga tggggctgca ggtgacccca aaaaggagaa gaaatctcta
      361 gactcagatg agagcgagga tgaagaagat gactaccagc aaaagcgcaa aggcgttgaa
      421 gggctcatcg acatcgagaa ccccaaccgg gtggcacaga caaccaaaaa ggtcacacaa
      481 ctggatctgg acgggccaaa ggagctttcg aggagagaac gagaagagat tgagaagcag
      541 aaggcaaaag agcgttacat gaaaatgcac ttggccggga agacagagca agccaaggct
      601 gacctggccc ggctggccat catccggaaa cagcgggagg aggctgcccg gaagaaggaa
      661 gaggaaagga aagcaaaaga cgatgccaca ttgtcaggaa aacgaatgca gtcactctcc
      721 ctgaataagt aagcggccgc tcgacctgca gccaagctta attagctgag cttggactcc
      781 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
      841 gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt ttcaggagct
      901 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
      961 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1021 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1081 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
     1141 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     1201 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1261 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     1321 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
     1381 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     1441 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     1501 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     1561 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg ggtaatgact
     1621 ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1681 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct ctagattacg
     1741 tgcagtcgat gataagctgt caaacatgag aattgtgcct aatgagtgag ctaacttaca
     1801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     1921 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     1981 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     2041 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     2101 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     2161 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     2221 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     2281 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     2341 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     2401 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     2461 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     2521 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     2581 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     2641 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     2701 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     2761 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     2821 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     2881 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     2941 cataccgcga aaggttttgc accattcgat ggtgtcggaa tttcgggcag cgttgggtcc
     3001 tggccacggg tgcgcatgat ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     3061 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3121 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     3181 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     3241 tactgagagt gcaccacatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3301 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3901 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5221 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5281 ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
