LOCUS       sequence-41896-e        5936 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 5936)
  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 5936)
  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-NOHMA
FEATURES             Location/Qualifiers
     source          1..5936
                     /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      1393..1487
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1531..2190
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1597..1616)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2255..2341
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2434..3516)
                     /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     3478..3497
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3517..3594)
                     /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(3678..3700)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3786..3926
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3841..3860
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3941..3958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4112..4700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4871..5662,5663..5731))
                     /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     5494..5513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5732..5836)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5904..5922
                     /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 ccactgccca gttgcagagg actcccatga gtgcactggt atttcccaat
      241 aagatatcaa ctgaacacca gtctttggtg ttagtgaaga ggcttctagc agtttcagta
      301 tcctgtatca cgtatttgag gggaatattc ccagaatgcg cttatggaac aagatatcta
      361 gatgatcttt gtgtcaaaat actgagagaa gataaaaatt gcccaggatc tacacagtta
      421 gtgaaatgga tgctaggatg ttatgatgct ttacagaaaa aatatgtata cacaaaccca
      481 gaagatcctc agacaatttc agaatgttac caattcaaat tcaaatacac caataatgga
      541 ccactcatgg acttcataag taaaaaccaa agcaacgaat ctagcatgtt gtctactgac
      601 accaagaaag caagcattct cctcattcgc aagatttata tcctaatgca aaatctgggg
      661 cctttaccta atgatgtttg tttgaccatg aaactttttt actatgatga agttacaccc
      721 ccagattacc agcctcccgg ttttaaggat ggtgattgtg aaggagttat atttgaaggg
      781 gaacctatgt atttaaatgt gggagaagtc tcaacacctt ttcacatctt caaagtaaaa
      841 gtgaccactg agagagaacg aatggaaaat attgactcaa ctatactatc accaaaacaa
      901 ataaaaacac catttcaaaa aatcctgagg gacaaagatg tagaagatga acaggagcat
      961 tatacaagtg atgatttgga cattgaaact aaaatggaag aacaggaaaa aaaccctgca
     1021 tcttctgaac ttgaagaacc aagtttagtt tgtgaggaag atgaaattat gaggtctaaa
     1081 gaaagtccag atctttctat ttctcattct caggttgagc agttagtcaa taaaacatct
     1141 gaacttgata tgtctgaaag caaaacaaga agtggaaaag tctttcagaa taaaatggca
     1201 aatggaaatc aaccagtaaa atcttccaaa gaaaatcgga agagaagtca acatgaatct
     1261 gggagaatag tcctccatca ctttgattct tctagtcaag agtcagtgcc aaaaaggaga
     1321 aagtttagtg aaccaaagga acatatatga gcggccgctc gacctgcagc caagcttaat
     1381 tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     1441 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagcttg
     1501 gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     1561 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     1621 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     1681 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     1741 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     1801 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     1861 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     1921 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     1981 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     2041 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     2101 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     2161 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
     2221 acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa
     2281 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2341 tccgccctct agattacgtg cagtcgatga taagctgtca aacatgagaa ttgtgcctaa
     2401 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2521 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     2581 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     2641 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     2701 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     2761 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     2821 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     2881 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     2941 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     3001 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     3061 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     3121 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     3181 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     3241 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     3301 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     3361 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     3421 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     3481 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     3541 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg tgtcggaatt
     3601 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct agagctgcct cgcgcgtttc
     3661 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     3721 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     3781 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     3841 cggcatcaga gcagattgta ctgagagtgc accacatgcg gtgtgaaata ccgcacagat
     3901 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     3961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     5881 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcac
//
