LOCUS       sequence-41857-e        5435 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 5435)
  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 5435)
  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-SUHW4
FEATURES             Location/Qualifiers
     source          1..5435
                     /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      892..986
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1030..1689
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1096..1115)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1754..1840
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1933..3015)
                     /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     2977..2996
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3016..3093)
                     /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(3177..3199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3285..3425
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3340..3359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3440..3457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3611..4199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3691..3710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4370..5161,5162..5230))
                     /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     4993..5012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5231..5335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5403..5421
                     /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 ctgaaagaga aacagaaaca tcaaattctg aaagtaaaca agataaagct
      241 gcttcttcaa aagaaaaaaa tggatgtaat gcaaattcat ttgaaggctc atcaacaaca
      301 aaaagtgaag aaagcataac agtttcagat aaggaaaatg aaacctgtct tgcagaccag
      361 gaaactggct caaaaaacat cgtcagttgt gattcaaata ttggtgcaga taaagtggaa
      421 aagaaaaaac aaatacaaca cgtttgtcag gaaatggagt tgaagatgtg ccagagttca
      481 gaaaacataa tcttatctga tcagattaaa gatcacaact ccagtgaagc cagattttct
      541 tcaaagaata ttaaggattt gcgattagca tcagataatg taagcattga tcagtttttg
      601 agaaaaagac atgaacctga atctgttagt tctgatgtta gcgagcaagg cagtattcat
      661 ttggaacctc tgactccatc cgaggtactt gagtatgaag ccacagagat tcttcagaaa
      721 ggtagtggtg atccttcagc caagactgat gaagtagtgt ctgatcaaac agatgacatt
      781 cctggaggaa ataaccctag cacaacagag gcaacagtag acctggaaga tgaaaaagaa
      841 agaagttgag cggccgctcg acctgcagcc aagcttaatt agctgagctt ggactcctgt
      901 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
      961 gccgggcgtt ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag
     1021 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1081 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1141 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1201 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     1261 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1321 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1381 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1441 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1501 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1561 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1621 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1681 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     1741 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1801 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gattacgtgc
     1861 agtcgatgat aagctgtcaa acatgagaat tgtgcctaat gagtgagcta acttacatta
     1921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     2041 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     2101 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     2161 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     2221 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     2281 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     2341 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     2401 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     2461 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     2521 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     2581 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     2641 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     2701 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     2761 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     2821 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     2881 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     2941 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     3001 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     3061 accgcgaaag gttttgcacc attcgatggt gtcggaattt cgggcagcgt tgggtcctgg
     3121 ccacgggtgc gcatgatcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3181 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3241 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3301 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     3361 tgagagtgca ccacatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3421 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4021 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4381 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4441 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4501 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4561 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4621 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4681 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4741 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4801 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4861 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5341 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5401 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
