LOCUS       sequence-250630-        2473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Research the effect of gene orientation of transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2473)
  AUTHORS   Yeung E, Dy AJ, Martin KB, Ng AH, Del Vecchio D, Beck JL, Collins 
            JJ, Murray RM
  TITLE     Biophysical Constraints Arising from Compositional Context in 
            Synthetic Gene Networks.
  JOURNAL   Cell Syst. 2017 Jul 26;5(1):11-24.e12. doi: 
            10.1016/j.cels.2017.06.001. Epub 2017 Jul 19.
  PUBMED    28734826
REFERENCE   2  (bases 1 to 2473)
  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"; doi: 
            "10.1016/j.cels.2017.06.001"; journalName: "Cell Syst"; date: 
            "2017-07-26- 26"; volume: "5"; issue: "1"; pages: "11-24.e12"
COMMENT     Sequence Label: pEYT1A
FEATURES             Location/Qualifiers
     source          1..2473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    44..62
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    69..87
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    319..335
                     /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)."
     rep_origin      complement(648..1236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(728..747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1407..2198,2199..2267))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2030..2049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2268..2372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2440..2458
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aattctggcg aatcctctga ccattccatt tccactaatc ttatccctat cagtgataga
       61 gattgacatc cctatcagtg atagagatac tgagcacagg atcccgactg gcgagagcca
      121 ggtaacgaat ggatcccaaa gcccgccgaa aggcgggctt ttctgttctt caccttaatc
      181 cattatacat aactcataca ttccaaacta atctaacttc ttccatcatt aaccacacct
      241 ctctatcatc acctattccc ctctatctcc ctcaaattct ttcacatatt atatcctaac
      301 actcatacac ccatacaatt gtgagcggat aacaagatac tgagcacagc ccggatagct
      361 cagtcggtag agcagcggcc ggacgcaact gaatgaaatg gtgaaggacg ggtccaggtg
      421 tggctgcttc ggcagtgcag cttgttgagt agagtgtgag ctccgtaact agtcgcgtcc
      481 ggccgcgggt ccagggttca agtccctgtt cgggcgccac aaagcccgcc gaaaggcggg
      541 cttttctgtt acccccttac aaaaccacag gataacgcag gaaagaacat ctgagcaaaa
      601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     2041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2401 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2461 tttcgtcttc aag
//
