LOCUS       sequence-14476-e        4027 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4027)
  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.
  PUBMED    15998469
REFERENCE   2  (bases 1 to 4027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb Cell
            Fact. 2005 Jul 5. 4():21."
COMMENT     Sequence Label: pQStrep2-ARPC3
FEATURES             Location/Qualifiers
     source          1..4027
                     /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             691..714
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             727..750
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      773..867
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             911..1570
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(977..996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1635..1721
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1769..1791)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1877..2017
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1932..1951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2032..2049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2203..2791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2283..2302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2962..3753,3754..3822))
                     /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     3585..3604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3823..3927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3995..4013
                     /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 ccggcttacc actcttctct catggatcct
      181 gataccaaac tcatcggaaa catggcactg ttgcctatca gaagtcaatt caaaggacct
      241 gcccccagag agacaaaaga tacagatatt gtggatgaag ccatctatta cttcaaggcc
      301 aatgtcttct tcaaaaacta tgaaattaag aatgaagctg ataggacctt gatatatata
      361 actctctaca tttctgaatg tctgaagaaa ctgcaaaagt gcaattccaa aagccaaggt
      421 gagaaagaaa tgtatacgct gggaatcact aattttccca ttcctggaga gcctggtttt
      481 ccacttaacg caatttatgc caaacctgca aacaaacagg aagatgaagt gatgagagcc
      541 tatttacaac agctaaggca agagactgga ctgagacttt gtgagaaagt tttcgaccct
      601 cagaatgata aacccagcaa gtggtggact tgctttgtga agagacagtt catgaacaag
      661 agtctttcag gacctggaca ggcggccgct tggagccacc cgcagttcga aaaataagcg
      721 gccgcttgga gccacccgca gttcgaaaaa taagcttaat tagctgagct tggactcctg
      781 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
      841 cgccgggcgt tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa
      901 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
      961 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1021 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1081 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     1141 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1201 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1261 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1321 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1381 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtctgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     1561 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct
     1621 ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1681 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgctc tagagctgcc
     1741 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     1801 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     1861 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     1921 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     1981 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     2041 tgactcgctg cgctcggtct gtcggctgcg gcgagcggta tcagctcact caaaggcggt
     2101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     2401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2641 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3001 ctgtctattt cgttcatcca tagctgcctg actccccgtc gtgtagataa ctacgatacg
     3061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3961 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4021 tcttcac
//
