LOCUS       sequence-41890-e        5225 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 5225)
  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 5225)
  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-C9orf58
FEATURES             Location/Qualifiers
     source          1..5225
                     /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      682..776
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             820..1479
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(886..905)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1544..1630
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1723..2805)
                     /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     2767..2786
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2806..2883)
                     /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(2967..2989)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3075..3215
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3130..3149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3230..3247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3401..3989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3481..3500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4160..4951,4952..5020))
                     /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     4783..4802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5021..5125)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5193..5211
                     /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 cagggatcct cgggcgagct cagcaacagg ttccaaggag ggaaggcgtt cggcttgctc
      241 aaagcccggc aggagaggag gctggccgag atcaaccggg agtttctgtg tgaccagaag
      301 tacagtgatg aagagaacct tccagaaaag ctcacagcct tcaaagagaa gtacatggag
      361 tttgacctga acaatgaagg cgagattgac ctgatgtctt taaagaggat gatggagaag
      421 cttggtgtcc ccaagaccca cctggagatg aagaagatga tctcagaggt gacaggaggg
      481 gtcagtgaca ctatatccta ccgagacttt gtgaacatga tgctggggaa acggtcggct
      541 gtcctcaagt tagtcatgat gtttgaagga aaagccaacg agagcagccc caagccagtt
      601 ggcccccctc cagagagaga cattgctagc ctgccctgag cggccgctcg acctgcagcc
      661 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      721 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
      781 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      841 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      901 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
      961 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1021 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1081 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1141 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1201 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1261 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1321 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1381 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1441 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     1501 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     1561 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1621 taggacaaat ccgccctcta gattacgtgc agtcgatgat aagctgtcaa acatgagaat
     1681 tgtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     1741 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1801 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     1861 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     1921 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     1981 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2041 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2101 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2161 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2221 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2281 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2341 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2401 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     2461 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     2521 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     2581 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     2641 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     2701 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     2761 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     2821 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     2881 gtcggaattt cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcta gagctgcctc
     2941 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3001 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3061 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccacatgcgg tgtgaaatac
     3181 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3241 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3301 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3421 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3481 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3541 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3841 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3901 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4021 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4081 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4141 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4201 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4261 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4321 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4381 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4441 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4501 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4561 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4621 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4681 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4741 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4801 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4861 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4921 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4981 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5041 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5101 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5161 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5221 ttcac
//
