LOCUS       sequence-41906-e        5213 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 5213)
  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 5213)
  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-C10orf83
FEATURES             Location/Qualifiers
     source          1..5213
                     /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      670..764
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             808..1467
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(874..893)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1532..1618
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1711..2793)
                     /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     2755..2774
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2794..2871)
                     /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(2955..2977)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3063..3203
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3118..3137
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3218..3235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4148..4939,4940..5008))
                     /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     4771..4790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5009..5113)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5181..5199
                     /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 cagggatcca ccctgacaaa aggttccttc acctactcca gtggggagga atatcgtggc
      241 gagtggaagg agggccgcag gcatggtttt ggtcaactga tgtttgcaga tggtggcacc
      301 tacctgggtc attttgagaa tgggctcttt aatggctttg gggtattgac cttctcagat
      361 ggttcaaggt atgaggggga gtttgcccag ggcaagttta atggcgtcgg agtcttcatt
      421 cgatatgaca acatgacctt tgagggggaa tttaaaaatg gcagagtaga tggttttggc
      481 ctgctgactt tccctgatgg ttctcatgga atcccccgca atgaaggtct ctttgagaac
      541 aacaagctgc tgcgacgtga gaagtgttct gccattgttc agcgggccca gagcgcctcc
      601 aagtcagcca gaaatctcac tgcctgagcg gccgctcgac ctgcagccaa gcttaattag
      661 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
      721 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
      781 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
      841 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
      901 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
      961 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1021 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1081 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1141 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1201 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1261 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1321 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1381 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1441 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1501 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1561 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1621 gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
     1681 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1741 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1801 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1861 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1921 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     1981 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     2041 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     2101 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     2161 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     2221 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     2281 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2341 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2401 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2461 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2521 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2581 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2641 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     2701 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     2761 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     2821 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cggaatttcg
     2881 ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt
     2941 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3001 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3061 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     3121 catcagagca gattgtactg agagtgcacc acatgcggtg tgaaataccg cacagatgcg
     3181 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5161 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
