LOCUS       sequence-41873-e        5411 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 5411)
  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 5411)
  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-RAB17
FEATURES             Location/Qualifiers
     source          1..5411
                     /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      868..962
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1006..1665
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1072..1091)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1730..1816
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1909..2991)
                     /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     2953..2972
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2992..3069)
                     /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(3153..3175)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3261..3401
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3316..3335
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3416..3433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3587..4175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3667..3686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4346..5137,5138..5206))
                     /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     4969..4988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5207..5311)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5379..5397
                     /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 cacaggcaca caggaccccc cagcccaggg ctgcccccag ccagccccgt
      241 gtgttcaagc tggttctcct gggaagtggc tccgtgggta agtccagctt ggctcttcgg
      301 tacgtgaaga acgacttcaa gagtatcctg cctacggtgg gctgtgcgtt cttcacaaag
      361 gtggtggatg tgggtgccac ctctctgaag cttgagatct gggacacagc tggccaggag
      421 aagtaccaca gcgtctgcca cctctacttc aggggtgcca acgctgcgct tctggtgtac
      481 gacatcacca ggaaggattc cttcctcaag gctcagcagt ggctgaagga cctggaggag
      541 gagctgcacc caggagaagt cctggtgatg ctggtgggca acaagacgga cctcagccag
      601 gagcgggagg tgaccttcca ggaagggaag gagtttgccg acagccagaa gttgccgttc
      661 atggaaactt cggccaaact gaaccaccag gtgtcggagg tgttcaatac agtggcccaa
      721 gagctactgc agagaagcga cgaggagggc caggctctac ggggggatgc agctgtggct
      781 ctgaacaagg ggcccgcgag gcaggccaaa tgctgcgccc actgagcggc cgctcgacct
      841 gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
      901 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
      961 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1021 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1081 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1141 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1201 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1261 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1321 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1381 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1441 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1501 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1561 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1621 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1681 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     1741 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1801 cctgagtagg acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat
     1861 gagaattgtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     1921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1981 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2041 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2101 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     2161 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     2221 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     2281 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     2341 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     2401 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     2461 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     2521 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     2581 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     2641 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     2701 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     2761 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     2821 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     2881 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     2941 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3001 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc
     3061 gatggtgtcg gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc
     3121 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3181 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3241 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3301 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccac atgcggtgtg
     3361 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4021 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5281 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5341 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5401 ttcgtcttca c
//
