LOCUS       sequence-14469-e        5261 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 5261)
  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 5261)
  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-SNX3
FEATURES             Location/Qualifiers
     source          1..5261
                     /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      718..812
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             856..1515
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(922..941)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1580..1666
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1759..2841)
                     /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     2803..2822
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2842..2919)
                     /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(3003..3025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3111..3251
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3166..3185
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3266..3283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3437..4025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3517..3536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4196..4987,4988..5056))
                     /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     4819..4838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5057..5161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5229..5247
                     /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 cggagaccgt ggctgacacc cggcggctga tcaccaagcc gcagaacctg
      241 aatgacgcct acggaccccc cagcaacttc ctcgagatcg atgtgagcaa cccgcaaacg
      301 gtgggggtcg gccggggccg cttcaccact tacgaaatca gggtcaagac aaatcttcct
      361 attttcaagc tgaaagaatc tactgttaga agaagataca gtgactttga atggctgcga
      421 agtgaattag aaagagagag caaggtcgta gttcccccgc tccctgggaa agcgtttttg
      481 cgtcagcttc cttttagagg agatgatgga atatttgatg acaattttat tgaggaaaga
      541 aaacaagggc tggagcagtt tataaacaag gtcgctggtc atcctctggc acagaacgaa
      601 cgttgtcttc acatgttttt acaagatgaa ataatagata aaagctatac tccatctaaa
      661 ataagacatg cctgagcggc cgctcgacct gcagccaagc ttaattagct gagcttggac
      721 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
      781 gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga
      841 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      901 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      961 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1021 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
     1081 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1141 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1201 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1261 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1321 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     1381 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1441 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1501 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     1561 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1621 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cctctagatt
     1681 acgtgcagtc gatgataagc tgtcaaacat gagaattgtg cctaatgagt gagctaactt
     1741 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1801 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     1861 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     1921 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     1981 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     2041 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     2101 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     2161 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     2221 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     2281 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     2341 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     2401 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     2461 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     2521 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     2581 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     2641 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     2701 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     2761 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     2821 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     2881 tgccataccg cgaaaggttt tgcaccattc gatggtgtcg gaatttcggg cagcgttggg
     2941 tcctggccac gggtgcgcat gatctagagc tgcctcgcgc gtttcggtga tgacggtgaa
     3001 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3061 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     3121 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     3181 ttgtactgag agtgcaccac atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3241 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3421 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3841 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4201 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4261 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4321 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4381 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4441 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4501 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4561 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4621 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4681 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4741 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4801 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4861 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4921 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4981 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5041 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5161 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5221 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
