LOCUS       sequence-41861-e        5366 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 5366)
  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 5366)
  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-SARA1
FEATURES             Location/Qualifiers
     source          1..5366
                     /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="ENLYFQA"
     terminator      823..917
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             961..1620
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1027..1046)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1685..1771
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1864..2946)
                     /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     2908..2927
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2947..3024)
                     /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(3108..3130)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3216..3356
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3271..3290
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3371..3388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3542..4130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3622..3641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4301..5092,5093..5161))
                     /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     4924..4943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5162..5266)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5334..5352
                     /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 caggcctctt tcatctttga gtggatctac aatggcttca gcagtgtgct ccagttccta
      241 ggactgtaca agaaatctgg aaaacttgta ttcttaggtt tggataatgc aggcaaaacc
      301 actcttcttc acatgctcaa agatgacaga ttgggccaac atgttccaac actacatccg
      361 acatcagaag agctaacaat tgctggaatg acctttacaa cttttgatct tggtgggcac
      421 gagcaagcac gtcgcgtttg gaaaaattat ctcccagcaa ttaatgggat tgtctttctg
      481 gtggactgtg cagatcattc tcgcctcgtg gaatccaaag ttgagcttaa tgctttaatg
      541 actgatgaaa caatatccaa tgtgccaatc cttatcttgg gtaacaaaat tgacagaaca
      601 gatgcaatca gtgaagaaaa actccgtgag atatttgggc tttatggaca gaccacagga
      661 aaggggaatg tgaccctgaa ggagctgaat gctcgcccca tggaagtgtt catgtgcagt
      721 gtgctcaaga ggcaaggtta cggcgagggt ttccgctggc tctcccagta tattgactga
      781 gcggccgctc gacctgcagc caagcttaat tagctgagct tggactcctg ttgatagatc
      841 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
      901 tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa
      961 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1021 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1081 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1141 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     1201 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1261 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1321 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1381 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1441 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1501 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     1561 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     1621 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag
     1681 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1741 gtcggtgaac gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga
     1801 taagctgtca aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg
     1861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     1981 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     2041 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     2101 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     2161 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     2221 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     2281 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     2341 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     2401 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     2461 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     2521 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     2581 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     2641 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     2701 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     2761 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     2821 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     2881 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     2941 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     3001 ggttttgcac cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg
     3061 cgcatgatct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3121 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3181 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3241 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3301 accacatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3361 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3421 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3481 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3541 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3601 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3661 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3721 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3781 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3841 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3901 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3961 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4021 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4081 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4141 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4201 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4261 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4321 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4381 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4441 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4501 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4561 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4621 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4681 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4741 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4801 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4861 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4921 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4981 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5041 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5101 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5161 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5221 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5281 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5341 gtatcacgag gccctttcgt cttcac
//
