LOCUS       sequence-14358-e        5846 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 5846)
  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 5846)
  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-LRPAP1
FEATURES             Location/Qualifiers
     source          1..5846
                     /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      1303..1397
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1441..2100
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1507..1526)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2165..2251
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2344..3426)
                     /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     3388..3407
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3427..3504)
                     /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(3588..3610)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3696..3836
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3751..3770
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3851..3868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4022..4610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4102..4121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4781..5572,5573..5641))
                     /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     5404..5423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5642..5746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5814..5832
                     /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 cgccgcggag ggtcaggtcg tttctgcgcg ggctcccggc gctgctactg
      241 ctgctgctct tcctcgggcc ctggcccgct gcgagccacg gcggcaagta ctcgcgggag
      301 aagaaccagc ccaagccgtc cccgaaacgc gagtccggag aggagttccg catggagaag
      361 ttgaaccagc tgtgggagaa ggcccagcga ctgcatcttc ctcccgtgag gctggccgag
      421 ctccacgctg atctgaagat acaggagagg gacgaactcg cctggaagaa actaaagctt
      481 gacggcttgg acgaagatgg ggagaaggaa gcgagactca tacgcaacct caatgtcatc
      541 ttggccaagt atggtctgga cggaaagaag gacgctcggc aggtgaccag caactccctc
      601 agtggcaccc aggaagacgg gctggatgac cccaggctgg aaaagctgtg gcacaaggcg
      661 aagacctctg ggaaattctc cggcgaagaa ctggacaagc tctggcggga gttcctgcat
      721 cacaaagaga aagttcacga gtacaacgtc ctgctggaga ccctgagcag gaccgaagaa
      781 atccacgaga acgtcattag cccctcggac ctgagcgaca tcaagggcag cgtcctgcac
      841 agcaggcaca cggagctgaa ggagaagctg cgcagcatca accagggcct ggaccgcctg
      901 cgcagggtca gccaccaggg ctacagcact gaggctgagt tcgaggagcc cagggtgatt
      961 gacctgtggg acctggcgca gtccgccaac ctcacggaca aggagctgga ggcgttccgg
     1021 gaggagctca agcacttcga agccaaaatc gagaagcaca accactacca gaagcagctg
     1081 gagattgcgc acgagaagct gaggcacgca gagagcgtgg gcgacggcga gcgtgtgagc
     1141 cgcagccgcg agaagcacgc cctgctggag gggcggacca aggagctggg ctacacggtg
     1201 aagaagcatc tgcaggacct gtccggcagg atctccagag ctcggcacaa cgaactctga
     1261 gcggccgctc gacctgcagc caagcttaat tagctgagct tggactcctg ttgatagatc
     1321 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     1381 tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa
     1441 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1501 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1561 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1621 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     1681 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1741 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1801 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1861 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1921 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1981 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     2041 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     2101 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag
     2161 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2221 gtcggtgaac gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga
     2281 taagctgtca aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg
     2341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     2461 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     2521 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     2581 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     2641 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     2701 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     2761 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     2821 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     2881 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     2941 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     3001 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     3061 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     3121 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     3181 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     3241 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     3301 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     3361 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     3421 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     3481 ggttttgcac cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg
     3541 cgcatgatct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3601 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3661 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3721 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3781 accacatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4441 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5641 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5701 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5761 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5821 gtatcacgag gccctttcgt cttcac
//
