LOCUS       sequence-41744-e        5363 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 5363)
  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 5363)
  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-HSPB8
FEATURES             Location/Qualifiers
     source          1..5363
                     /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      820..914
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             958..1617
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1024..1043)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1682..1768
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1861..2943)
                     /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     2905..2924
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2944..3021)
                     /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(3105..3127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3213..3353
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3268..3287
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3368..3385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3539..4127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3619..3638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4298..5089,5090..5158))
                     /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     4921..4940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5159..5263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5331..5349
                     /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 cagggatctg ctgacggtca gatgcccttc tcctgccact acccaagccg cctgcgccga
      241 gaccccttcc gggactctcc cctctcctct cgcctgctgg atgatggctt tggcatggac
      301 cccttcccag acgacttgac agcctcttgg cccgactggg ctctgcctcg tctctcctcc
      361 gcctggccag gcaccctaag gtcgggcatg gtgccccggg gccccactgc caccgccagg
      421 tttggggtgc ctgccgaggg caggaccccc ccacccttcc ctggggagcc ctggaaagtg
      481 tgtgtgaatg tgcacagctt caagccagag gagttgatgg tgaagaccaa agatggatac
      541 gtggaggtgt ctggcaaaca tgaagagaaa cagcaagaag gtggcattgt ttctaagaac
      601 ttcacaaaga aaatccagct tcctgcagag gtggatcctg tgacagtatt tgcctcactt
      661 tccccagagg gtctgctgat catcgaagct ccccaggtcc ctccttactc aacatttgga
      721 gagagcagtt tcaacaacga gcttccccag gacagccagg aagtcacctg tacctgagcg
      781 gccgctcgac ctgcagccaa gcttaattag ctgagcttgg actcctgttg atagatccag
      841 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
      901 ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg
      961 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1021 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1081 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1141 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     1201 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1261 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1321 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1381 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1441 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1501 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1561 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     1621 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     1681 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1741 ggtgaacgct ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa
     1801 gctgtcaaac atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     1861 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1921 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     1981 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     2041 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     2101 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     2161 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     2221 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     2281 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     2341 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     2401 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     2461 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     2521 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     2581 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     2641 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     2701 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     2761 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     2821 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     2881 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     2941 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     3001 tttgcaccat tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc
     3061 atgatctaga gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     3121 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3181 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     3241 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     3301 acatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     3361 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3421 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3961 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5281 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5341 tcacgaggcc ctttcgtctt cac
//
