LOCUS       sequence-41902-e        5891 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 5891)
  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 5891)
  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-MGC20398
FEATURES             Location/Qualifiers
     source          1..5891
                     /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      1348..1442
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1486..2145
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1552..1571)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2210..2296
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2389..3471)
                     /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     3433..3452
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3472..3549)
                     /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(3633..3655)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3741..3881
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3796..3815
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3896..3913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4067..4655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4147..4166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4826..5617,5618..5686))
                     /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     5449..5468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5687..5791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5859..5877
                     /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 cgtcctccgc ctccgcccgg actccggcag ggaagcgagt gataaatcag
      241 gaagaattgc ggcggttaat gaaggagaag cagcgtctga gcaccagtcg gaaacggata
      301 gaatctccat tcgcgaagta caaccgtttg gggcagctga gttgtgccct gtgtaacact
      361 ccggttaaga gcgagctcct gtggcagact cacgtcctgg gaaagcagca ccgagagaaa
      421 gtggccgagc tgaaaggcgc gaaggaagcc agccagggtt cgtccgccag ttcagcgcct
      481 cagtccgtca agaggaaagc gccggacgca gacgaccaag atgtcaagag agcgaaggcc
      541 accttggtgc ctcaggtaca gccctccaca tctgcgtgga ccaccaactt tgacaaaata
      601 ggaaaggagt tcattagagc gactcccagt aagccttcag gactcagttt actccccgat
      661 tatgaagatg aggaggagga ggaagaggag gaggaaggag atggagaaag aaaaaggggg
      721 gacgccagca agccgctctc cgacgcacag ggcaaggagc actcagtttc ctcttcacgg
      781 gaggtaacaa gtagtgtgct gccaaacgat ttctttagta ctaatcctcc caaggccccc
      841 ataattcctc attcagggtc aattgagaaa gcagaaatac atgaaaaagt ggtggaaagg
      901 agagaaaaca ccgcggaagc gttaccggaa ggtttttttg acgaccctga ggtagatgca
      961 agagtacgaa aggttgatgc tccaaaagat cagatggaca aagagtggga cgaattccaa
     1021 aaagccatga ggcaggtcaa cactatttcc gaagccatag ttgccgaaga ggatgaggag
     1081 ggacggttgg accgccagat tggggagatc gatgagcaga tagagtgtta ccgacgggtg
     1141 gaaaagctac ggaatcgcca ggatgaaata aaaaataaac ttaaagaaat cctgaccata
     1201 aaagaactgc agaaaaagga agaagagaat gctgacagcg atgatgaggg ggaactacag
     1261 gatttgttgt ctcaggattg gagggtgaaa ggggcattgt tataggcggc cgctcgacct
     1321 gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
     1381 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     1441 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1501 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1561 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1621 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1681 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1741 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1801 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1861 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1921 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1981 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2041 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2101 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     2161 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     2221 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2281 cctgagtagg acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat
     2341 gagaattgtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2401 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2461 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2521 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2581 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     2641 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     2701 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     2761 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     2821 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     2881 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     2941 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     3001 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     3061 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     3121 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     3181 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     3241 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     3301 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     3361 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     3421 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3481 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc
     3541 gatggtgtcg gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc
     3601 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3661 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3721 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3781 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccac atgcggtgtg
     3841 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3901 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3961 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4021 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4081 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4141 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4201 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4261 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4321 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4381 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4441 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4501 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4561 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4621 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4681 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5701 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5761 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5821 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5881 ttcgtcttca c
//
