LOCUS       sequence-244811-        3384 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 3384)
  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 3384)
  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: pEYD1A
FEATURES             Location/Qualifiers
     source          1..3384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..40
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    47..65
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             106..783
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     protein_bind    1186..1202
                     /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    1209..1225
                     /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(1538..2126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1618..1637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2297..3088,3089..3157))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2920..2939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3158..3262)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3330..3348
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttccatttc cactaatctt atccctatca gtgatagaga ttgacatccc tatcagtgat
       61 agagatactg agcacggatc ccgactttaa gaaggagata tacatatggc gagtagcgaa
      121 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
      181 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
      241 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
      301 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
      361 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
      421 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
      481 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
      541 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
      601 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
      661 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
      721 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
      781 taagccaggt aacgaatgga tcccaaagcc cgccgaaagg cgggcttttc tgttcttcac
      841 cttaatccat tatacataac tcaataccta cccattcctc ttaacatcaa tctaattata
      901 tatcattatt accctacact taaaccccat ctcaatactt cccatcaaat ccaacctaac
      961 tcatacattc caaactaatc taacttcttc catcattaac cacacctctc tatcatcacc
     1021 tattcccctc tatctccctc aaattctttc acatattaaa aacttactct tcctcctttc
     1081 ctccttaaaa tactaaataa ctcttaaata ccctaatctt tcacttctat tacccaaata
     1141 tcttatcatc accaacctta tcctaacact catacaccca tacaattgtg agcggataac
     1201 aattgacatt gtgagcggat aacaagatac tgagcacagc ccggatagct cagtcggtag
     1261 agcagcggcc ggacgcaact gaatgaaatg gtgaaggacg ggtccaggtg tggctgcttc
     1321 ggcagtgcag cttgttgagt agagtgtgag ctccgtaact agtcgcgtcc ggccgcgggt
     1381 ccagggttca agtccctgtt cgggcgccac aaagcccgcc gaaaggcggg cttttctgtt
     1441 acccccttac aaaaccacag gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1501 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1561 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1621 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1681 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     1741 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1801 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1861 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1921 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     1981 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2041 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2101 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2161 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2221 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2281 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2341 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2401 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2461 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2521 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2581 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2641 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2701 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2761 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2821 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2881 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2941 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3001 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3061 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3121 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3181 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3241 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3301 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     3361 aagaattctg gcgaatcctc tgac
//
