LOCUS       sequence-41855-e        6395 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 6395)
  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 6395)
  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-MPP6
FEATURES             Location/Qualifiers
     source          1..6395
                     /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      1852..1946
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1990..2649
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2056..2075)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2714..2800
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2893..3975)
                     /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     3937..3956
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3976..4053)
                     /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(4137..4159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4245..4385
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4300..4319
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4400..4417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4571..5159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4651..4670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5330..6121,6122..6190))
                     /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     5953..5972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6191..6295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6363..6381
                     /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 cagggatccc agcaagtctt ggaaaacctt acggagctgc cctcgtctac tggagcagaa
      241 gaaatagacc taattttcct caagggaatt atggagaatc ctattgtaaa atcacttgct
      301 aaggctcatg agaggctaga agattccaaa ctagaagctg tcagtgacaa taacttggaa
      361 ttagtcaatg aaattcttga agacatcact cctctaataa atgtggatga aaatgtggca
      421 gaattggttg gtatactcaa agaacctcac ttccagtcac tgttggaggc ccatgatatt
      481 gtggcatcaa agtgttatga ttcacctcca tcaagcccag aaatgaataa ttcttctatc
      541 aataatcagt tattaccagt agatgccatt cgtattcttg gtattcacaa aagagctggg
      601 gaaccactgg gtgtgacatt tagggttgaa aataatgatc tggtaattgc ccgaatcctc
      661 catgggggaa tgatagatcg acaaggtcta cttcatgtgg gagatataat taaagaagtc
      721 aatggccatg aggttggaaa taatccaaag gaattacaag aattactgaa aaatattagt
      781 ggaagtgtca ccctaaaaat cttaccaagt tatagagata ccattactcc tcaacaggta
      841 tttgtgaagt gtcattttga ttataatcca tacaatgaca acctaatacc ttgcaaagaa
      901 gcaggattga agttttccaa aggagaaatt cttcagattg taaatagaga agatccaaat
      961 tggtggcagg ctagccatgt aaaagaggga ggaagcgctg gtctcattcc aagccagttc
     1021 ctggaagaga agagaaaggc atttgttaga agagactggg acaattcagg acctttttgc
     1081 ggaactataa gtagcaaaaa aaagaaaaag atgatgtatc tcacaaccag aaatgcagaa
     1141 tttgatcgtc atgaaatcca gatatatgag gaggtagcca aaatgcctcc cttccagaga
     1201 aaaacattag tattgatagg agctcaaggt gtaggccgaa gaagcttgaa aaacaggttc
     1261 atagtattga atcccactag atttggaact acggtgccat ttacttcacg gaaaccaagg
     1321 gaagatgaaa aagatggcca ggcatataag tttgtgtcac gatctgagat ggaagcagat
     1381 attaaagctg gaaagtattt ggaacatggg gaatatgaag gaaatctcta tggaaccaaa
     1441 attgattcta ttcttgaggt tgtccaaact ggacggactt gcattctgga tgtcaaccca
     1501 caagcactga aagtattgag gacatcagag tttatgccct atgtggtatt tattgcggct
     1561 ccggagctag agacgttacg tgccatgcac aaggctgtgg tggatgcagg aatcactacc
     1621 aagcttctga ccgactctga cttgaagaaa acagtggatg aaagtgcacg gattcagaga
     1681 gcatacaacc actattttga tttgatcatc ataaatgata atctagacaa agcctttgaa
     1741 aaactgcaaa ctgccataga gaaactgaga atggaaccac agtgggtccc aatcagctgg
     1801 gtttactgag cggccgctcg acctgcagcc aagcttaatt agctgagctt ggactcctgt
     1861 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1921 gccgggcgtt ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag
     1981 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2041 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2101 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2161 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     2221 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2281 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2341 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2401 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2461 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2521 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2581 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2641 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     2701 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2761 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gattacgtgc
     2821 agtcgatgat aagctgtcaa acatgagaat tgtgcctaat gagtgagcta acttacatta
     2881 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2941 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     3001 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     3061 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     3121 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     3181 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     3241 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     3301 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     3361 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     3421 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     3481 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     3541 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     3601 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     3661 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     3721 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     3781 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     3841 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     3901 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     3961 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     4021 accgcgaaag gttttgcacc attcgatggt gtcggaattt cgggcagcgt tgggtcctgg
     4081 ccacgggtgc gcatgatcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4141 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4201 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4261 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4321 tgagagtgca ccacatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4381 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4981 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6301 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6361 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
