LOCUS       sequence-14364-e        6041 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 6041)
  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 6041)
  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-PSMD11
FEATURES             Location/Qualifiers
     source          1..6041
                     /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      1498..1592
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1636..2295
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1702..1721)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2360..2446
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2539..3621)
                     /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     3583..3602
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3622..3699)
                     /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(3783..3805)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3891..4031
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3946..3965
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4046..4063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4217..4805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4297..4316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4976..5767,5768..5836))
                     /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     5599..5618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5837..5941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6009..6027
                     /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 cggcggcggc ggtggtggag ttccagagag cccagtctct actcagcacc
      241 gaccgggagg cctccatcga catcctccac tccatcgtga agcgtgacat tcaggaaaac
      301 gatgaagagg cagtgcaagt caaagagcag agcatcctgg aactgggatc tctcctggca
      361 aagactggac aagctgcaga gcttggagga ctcctgaagt atgtacgacc cttcttgaat
      421 tccatcagca aggctaaagc agctcgcctg gtccgatctc ttcttgatct gtttcttgat
      481 atggaagcag ctacagggca ggaggtcgag ctgtgtttag agtgcatcga atgggccaag
      541 tcagagaaaa gaactttctt acgccaagct ttggaggcaa gactggtgtc tttgtacttt
      601 gataccaaga ggtaccagga agcattgcat ttgggttctc agctgctgcg ggagttgaaa
      661 aagatggacg acaaagctct tttggtggaa gtacagcttt tagaaagcaa aacataccat
      721 gccctgagca acctgccgaa agcccgagct gccttaactt ctgctcgaac cacagcaaat
      781 gccatctact gcccccctaa attgcaggcc accttggaca tgcagtcggg tattatccat
      841 gcagcagaag agaaggactg gaaaactgcg tactcatact tctatgaggc atttgagggt
      901 tatgactcca tcgacagccc caaggccatc acatctctga agtacatgtt gctgtgcaaa
      961 atcatgctca acaccccaga agatgtccag gctttggtga gcgggaagct tgcacttcgg
     1021 tatgcaggga ggcagacaga agcattaaaa tgcgtggctc aggctagcaa gaacagatca
     1081 ctggcagatt ttgaaaaggc tctgacagat taccgggcag agctccggga tgacccaatc
     1141 atcagcacac acttggccaa gttgtatgat aacttactag aacagaatct gatccgagtc
     1201 attgagcctt tttccagagt acagattgaa cacatatcta gtctcatcaa actctccaag
     1261 gccgacgtgg aaaggaaatt atcacagatg attcttgaca agaaatttca tgggattttg
     1321 gaccaggggg agggtgtcct gattattttc gatgaacccc cagtagataa aacttacgaa
     1381 gctgctctgg aaacaattca gaacatgagc aaagtagtgg attccctcta caacaaagcc
     1441 aagaaactga cataggcggc cgctcgacct gcagccaagc ttaattagct gagcttggac
     1501 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     1561 gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga
     1621 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1681 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1741 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1801 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
     1861 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1921 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1981 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     2041 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     2101 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     2161 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     2221 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     2281 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     2341 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2401 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cctctagatt
     2461 acgtgcagtc gatgataagc tgtcaaacat gagaattgtg cctaatgagt gagctaactt
     2521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     2641 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     2701 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     2761 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     2821 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     2881 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     2941 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     3001 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     3061 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     3121 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     3181 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     3241 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     3301 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     3361 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     3421 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     3481 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     3541 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     3601 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     3661 tgccataccg cgaaaggttt tgcaccattc gatggtgtcg gaatttcggg cagcgttggg
     3721 tcctggccac gggtgcgcat gatctagagc tgcctcgcgc gtttcggtga tgacggtgaa
     3781 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3841 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     3901 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     3961 ttgtactgag agtgcaccac atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4021 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4081 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4141 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4621 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4681 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4801 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4861 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4921 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5041 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5161 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5401 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5641 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5821 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5881 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5941 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6001 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
