LOCUS       sequence-14466-e        4363 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 4363)
  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 4363)
  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-PIR
FEATURES             Location/Qualifiers
     source          1..4363
                     /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             1027..1050
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1063..1086
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      1109..1203
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1247..1906
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1313..1332)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1971..2057
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2105..2127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2213..2353
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2268..2287
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2368..2385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2539..3127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2619..2638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3298..4089,4090..4158))
                     /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     3921..3940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4159..4263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4331..4349
                     /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 tcacggatct gggtcctcca agaaagttac tctctcagtg
      181 ctcagccggg agcagtcgga aggggttgga gcgagggtcc ggagaagcat tggcagaccc
      241 gagttaaaaa atctggatcc gtttttactg tttgatgaat ttaaaggagg tagaccagga
      301 ggatttcctg atcatccaca tcgaggtttt gaaacagtat cctacctcct ggaagggggc
      361 agcatggccc atgaagactt ctgtggacac actggtaaaa tgaacccagg agatttgcag
      421 tggatgactg cgggccgggg cattctgcac gctgagatgc cttgctcaga ggagccagcc
      481 catggcctac aactgtgggt taatttgagg agctcagaga agatggtgga gcctcagtac
      541 caggaactga aaagtgaaga aatccctaaa cccagtaagg atggtgtgac agttgctgtc
      601 atttctggag aagccctggg aataaagtcc aaggtttaca ctcgcacacc aaccttatat
      661 ttggacttca aattggaccc aggagccaaa cattcccaac ctatccctaa agggtggaca
      721 agcttcattt acacgatatc tggagatgtg tatattgggc ccgatgatgc acaacaaaaa
      781 atagaacctc atcacacagc agtgcttgga gaaggtgaca gtgtccaggt ggagaacaag
      841 gatcccaaga gaagccactt tgtcttaatt gctggggagc cattaagaga accagttatc
      901 caacatggtc catttgtgat gaacaccaat gaagagattt ctcaagctat tcttgatttc
      961 agaaacgcaa aaaatgggtt tgaaagggcc aaaacctgga aatcaaagat tgggaacgcg
     1021 gccgcttgga gccacccgca gttcgaaaaa taagcggccg cttggagcca cccgcagttc
     1081 gaaaaataag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
     1141 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     1201 aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1261 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1321 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1381 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1441 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
     1501 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1561 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1621 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1681 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1741 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     1801 gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct
     1861 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     1921 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
     1981 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2041 tcctgagtag gacaaatccg ccgctctaga gctgcctcgc gcgtttcggt gatgacggtg
     2101 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2161 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     2221 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     2281 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2341 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtctgtcg
     2401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2701 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     2761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2941 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagc
     3361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4261 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4321 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
