LOCUS       sequence-14480-e        3970 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 3970)
  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 3970)
  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-GADD45G
FEATURES             Location/Qualifiers
     source          1..3970
                     /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             634..657
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             670..693
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      716..810
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             854..1513
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(920..939)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1578..1664
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1712..1734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1820..1960
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1875..1894
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1975..1992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2146..2734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2226..2245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2905..3696,3697..3765))
                     /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     3528..3547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3766..3870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3938..3956
                     /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 actctggaag aagtccgcgg ccaggacaca
      181 gttccggaaa gcacagccag gatgcagggt gccgggaaag cgctgcatga gttgctgctg
      241 tcggcgcagc gtcagggctg cctcactgcc ggcgtctacg agtcagccaa agtcttgaac
      301 gtggaccccg acaatgtgac cttctgtgtg ctggctgcgg gtgaggagga cgagggcgac
      361 atcgcgctgc agatccattt tacgctgatc caggctttct gctgcgagaa cgacatcgac
      421 atagtgcgcg tgggcgatgt gcagcggctg gcggctatcg tgggcgccgg cgaggaggcg
      481 ggtgcgccgg gcgacctgca ctgcatcctc atttcgaacc ccaacgagga cgcctggaag
      541 gatcccgcct tggagaagct cagcctgttt tgcgaggaga gccgcagcgt taacgactgg
      601 gtgcccagca tcaccctccc cgaggcggcc gcttggagcc acccgcagtt cgaaaaataa
      661 gcggccgctt ggagccaccc gcagttcgaa aaataagctt aattagctga gcttggactc
      721 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
      781 tgccgccggg cgttttttat tggtgagaat ccaagctagc ttggcgagat tttcaggagc
      841 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
      901 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
      961 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     1021 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc
     1081 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     1141 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     1201 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1261 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     1321 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     1381 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtctg
     1441 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     1501 gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac
     1561 tctctagctt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1621 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ctctagagct
     1681 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1741 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1801 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     1861 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     1921 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     1981 cactgactcg ctgcgctcgg tctgtcggct gcggcgagcg gtatcagctc actcaaaggc
     2041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2341 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2941 gatctgtcta tttcgttcat ccatagctgc ctgactcccc gtcgtgtaga taactacgat
     3001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3901 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3961 tcgtcttcac
//
