LOCUS       sequence-41897-e        5405 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 5405)
  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 5405)
  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-CIP29
FEATURES             Location/Qualifiers
     source          1..5405
                     /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      862..956
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1000..1659
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1066..1085)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1724..1810
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1903..2985)
                     /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     2947..2966
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2986..3063)
                     /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(3147..3169)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3255..3395
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3310..3329
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3410..3427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3581..4169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3661..3680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4340..5131,5132..5200))
                     /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     4963..4982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5201..5305)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5373..5391
                     /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 cgaccgagac ggtggagctc cataagctaa agcttgccga actaaagcaa
      241 gaatgtcttg ctcgtggttt ggagaccaag ggaataaagc aagatcttat ccacagactc
      301 caggcatatc ttgaagaaca tgctgaagag gaggcaaatg aagaagatgt actgggagat
      361 gaaacagagg aagaagaaac aaagcccatt gagctccctg tcaaagagga agaaccccct
      421 gaaaaaactg ttgatgtggc agcagagaag aaagtggtga aaattacatc tgaaatacca
      481 cagactgaga gaatgcagaa gagggctgaa cgattcaatg tacctgtgag cttggagagt
      541 aagaaagctg ctcgggcagc taggtttggg atttcttcag ttccaacaaa aggtctgtca
      601 tctgataaca aacctatggt taacttggat aagctgaagg aaagagctca aagatttggt
      661 ttgaatgtct cttcaatctc cagaaagtct gaagatgatg agaaactgaa aaagaggaag
      721 gagcgatttg ggattgtcac aagttcagct ggaactggaa ccacagagga tacagaggca
      781 aagaagagga aaagagcaga gcgctttggg attgcctgag cggccgctcg acctgcagcc
      841 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      901 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
      961 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1021 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1081 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1141 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1201 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1261 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1321 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1381 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1441 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1501 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1561 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1621 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     1681 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     1741 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1801 taggacaaat ccgccctcta gattacgtgc agtcgatgat aagctgtcaa acatgagaat
     1861 tgtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     1921 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1981 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2041 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2101 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2161 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2221 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2281 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2341 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2401 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2461 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2521 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2581 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     2641 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     2701 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     2761 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     2821 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     2881 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     2941 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3001 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     3061 gtcggaattt cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcta gagctgcctc
     3121 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3181 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3241 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3301 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccacatgcgg tgtgaaatac
     3361 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4021 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5341 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5401 ttcac
//
