LOCUS       sequence-14474-e        5765 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 5765)
  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 5765)
  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-PHYHIP
FEATURES             Location/Qualifiers
     source          1..5765
                     /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      1222..1316
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1360..2019
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1426..1445)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2084..2170
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2263..3345)
                     /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     3307..3326
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3346..3423)
                     /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(3507..3529)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3615..3755
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3670..3689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3770..3787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3941..4529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4021..4040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4700..5491,5492..5560))
                     /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     5323..5342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5561..5665)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5733..5751
                     /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 agctgctgtc cacgccccac agcattgaga tcaacaacat cacctgcgac
      241 tccttccgca tctcctgggc catggaggac agtgacctgg agagggtcac ccattacttc
      301 attgacctta acaagaagga gaataagaat tccaacaagt tcaagcaccg ggacgtcccc
      361 accaagctcg tggccaaggc agtgccgctg cccatgacgg tgagaggcca ctggttcctg
      421 agcccccgca cggagtacag tgtggccgtg cagacggcag tgaagcagag cgatggggag
      481 tacctggtgt ccggctggag cgagacggtg gagttctgca ctggggatta tgccaaggag
      541 cacctggctc agcttcagga gaaagctgag cagatcgcag gccgcatgct ccgcttctcc
      601 gtcttctacc gcaaccatca caaggagtac ttccagcatg ccaggaccca ctgcgggaac
      661 atgctgcagc cttacctgaa ggacaacagc ggcagccacg gctcccccac cagcggtatg
      721 ctccacgggg tcttcttcag ctgcaacacg gagttcaaca cgggccagcc cccgcaggac
      781 tccccctacg gccgctggcg cttccagatc ccagctcagc gcctcttcaa tcccagcacc
      841 aacctctact ttgcggactt ctactgcatg tacacggcct accactacgc catcctggtg
      901 ctggcgccca aaggctcccc gggggaccgc ttctgccgcg accgcctgcc cctcctggac
      961 attgcttgca acaagttcct gacctgcagc gtggaggatg gggagctggt cttccgccac
     1021 gcccaggacc tcatcctgga gatcatctac actgagcccg tcgacctgtc cctgggcacc
     1081 ctgggggaga tcagtgggca ccagctcatg agtctgtcta ctgccgatgc caagaaggac
     1141 cccagctgca agacctgcaa catcagcgtg ggccgctgag cggccgctcg acctgcagcc
     1201 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1261 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
     1321 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1381 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1441 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1501 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1561 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1621 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1681 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1741 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1801 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1861 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1921 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1981 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     2041 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     2101 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     2161 taggacaaat ccgccctcta gattacgtgc agtcgatgat aagctgtcaa acatgagaat
     2221 tgtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     2281 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2341 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2401 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2461 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2521 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2581 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2641 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2701 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2761 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2821 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2881 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2941 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     3001 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     3061 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     3121 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     3181 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     3241 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     3301 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3361 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     3421 gtcggaattt cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcta gagctgcctc
     3481 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3541 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3601 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3661 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccacatgcgg tgtgaaatac
     3721 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3781 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3841 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4381 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5761 ttcac
//
