LOCUS       sequence-14487-e        4168 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 4168)
  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 4168)
  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: pQStrep2-PREI3
FEATURES             Location/Qualifiers
     source          1..4168
                     /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             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             832..855
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             868..891
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      914..1008
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1052..1711
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1118..1137)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1776..1862
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1910..1932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2018..2158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2073..2092
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2173..2190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3103..3894,3895..3963))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3726..3745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3964..4068)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4136..4154
                     /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 aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc gtcatggcgg aggggacggc agtgctgagg
      181 cggaacaggc cgggcaccaa ggcgcaggat ttctataatt ggcctgatga atcctttgat
      241 gaaatggaca gtacactagc tgttcaacag tatattcaac agaacataag agcagattgc
      301 tccaatattg acaaaattct tgaaccacct gaaggccaag atgaaggtgt gtggaagtat
      361 gaacatttaa ggcagttctg ccttgagcta aatggacttg ctgtcaaact tcagagtgaa
      421 tgccatccag atacttgcac tcaaatgaca gcaactgaac aatggatttt tctttgtgca
      481 gctcataaaa ctccaaaaga gtgtcctgct atagactata ctagacacac acttgatggt
      541 gctgcatgtc ttctgaatag caataaatat tttcccagca gggttagcat aaaggaatca
      601 tctgtagcga aactaggatc agtatgccgt aggatttaca gaatattttc acatgcttat
      661 tttcatcatc ggcagatatt tgatgaatat gaaaatgaaa catttttgtg tcatcggttt
      721 actaagtttg tgatgaaata caatttgatg tccaaggata acctgattgt accaatttta
      781 gaagaggaag tacagaattc agtttctggg gaaagtgaag cagcggccgc ttggagccac
      841 ccgcagttcg aaaaataagc ggccgcttgg agccacccgc agttcgaaaa ataagcttaa
      901 ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
      961 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagctt
     1021 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     1081 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     1141 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     1201 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     1261 tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     1321 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     1381 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1441 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     1501 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1561 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     1621 ggttcatcat gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     1681 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     1741 aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga
     1801 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1861 atccgccgct ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1921 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1981 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     2041 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     2101 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc tgtcggctgc ggcgagcggt
     2221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagctgcct gactccccgt
     3181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4081 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4141 gcgtatcacg aggccctttc gtcttcac
//
