LOCUS       sequence-41892-e        6215 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 6215)
  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 6215)
  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-SESN2
FEATURES             Location/Qualifiers
     source          1..6215
                     /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      1672..1766
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1810..2469
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1876..1895)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2534..2620
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2713..3795)
                     /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     3757..3776
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3796..3873)
                     /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(3957..3979)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4065..4205
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4120..4139
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4220..4237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4471..4490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5150..5941,5942..6010))
                     /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     5773..5792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6011..6115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6183..6201
                     /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 tcgtggcgga ctccgagtgc cgcgcagagc tcaaggacta cctgcggttc
      241 gccccgggcg gcgtcggcga ctcgggcccc ggagaggagc agagggagag ccgggctcgg
      301 cgaggccctc gagggcccag cgccttcatc cccgtggagg aggtccttcg ggagggggct
      361 gagagcctcg agcagcacct ggggctggag gcactgatgt cctctgggcg agtagacaac
      421 ctggcagtgg tgatgggcct gcaccctgac tactttacca gcttctggcg cctgcactac
      481 ctgctgctgc acacggatgg tcccttggcc agctcctggc gccactacat tgccatcatg
      541 gctgccgccc gccatcagtg ttcttacctg gtaggctccc acatggccga gtttctgcag
      601 actggtggtg accctgagtg gctgctgggc ctccaccggg cccccgagaa gctgcgcaaa
      661 ctcagcgaga tcaacaagtt gctggcgcat cggccatggc tcatcaccaa ggaacacatc
      721 caggccttgc tgaagaccgg cgagcacact tggtccctgg ccgagctcat tcaggctctg
      781 gtcctgctca cccactgcca ctcgctctcc tccttcgtgt ttggctgtgg catcctccct
      841 gagggggatg cagatggcag ccctgccccc caggcaccta caccccctag tgaacagagc
      901 agccccccaa gcagggaccc gttgaacaac tctgggggct ttgagtctgc ccgcgacgtg
      961 gaggcgctga tggagcgcat gcagcagctg caggagagcc tgctgcggga tgaggggacg
     1021 tcccaggagg agatggagag ccgctttgag ctggagaagt cagagagcct gctggtgacc
     1081 ccctcagctg acatcctgga gccctctcca cacccagaca tgctgtgctt tgtggaagac
     1141 cctactttcg gatatgagga cttcactcgg agaggggctc aggcaccccc taccttccgg
     1201 gcccaggatt atacctggga agaccatggc tactcgctga tccagcggct ttaccctgag
     1261 ggtgggcagc tgctggatga gaagttccag gcagcctata gcctcaccta caataccatc
     1321 gccatgcaca gtggtgtgga cacctccgtg ctccgcaggg ccatctggaa ctatatccac
     1381 tgcgtctttg gcatcagata tgatgactat gattatgggg aggtgaacca gctcctggag
     1441 cggaacctca aggtctatat caagacagtg gcctgctacc cagagaagac cacccgaaga
     1501 atgtacaacc tcttctggag gcacttccgc cactcagaga aggtccacgt gaacttgctg
     1561 ctcctggagg cgcgcatgca agccgctctg ctgtacgccc tccgtgccat cacccgctac
     1621 atgacctgag cggccgctcg acctgcagcc aagcttaatt agctgagctt ggactcctgt
     1681 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1741 gccgggcgtt ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag
     1801 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1861 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1921 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1981 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     2041 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2101 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2161 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2221 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2281 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2341 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2401 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2461 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     2521 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2581 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gattacgtgc
     2641 agtcgatgat aagctgtcaa acatgagaat tgtgcctaat gagtgagcta acttacatta
     2701 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2761 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     2821 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     2881 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     2941 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     3001 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     3061 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     3121 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     3181 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     3241 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     3301 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     3361 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     3421 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     3481 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     3541 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     3601 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     3661 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     3721 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     3781 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     3841 accgcgaaag gttttgcacc attcgatggt gtcggaattt cgggcagcgt tgggtcctgg
     3901 ccacgggtgc gcatgatcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3961 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4021 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4081 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4141 tgagagtgca ccacatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4201 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4801 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6121 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6181 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
