LOCUS       sequence-258356-        3806 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 3806)
  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 3806)
  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: pEYRC-C1A
FEATURES             Location/Qualifiers
     source          1..3806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..57
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    64..82
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             123..800
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1075..1096)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1424..1443
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1683..1704
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1839..1855
                     /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(1976..2564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2056..2075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2735..3526,3527..3595))
                     /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     3358..3377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3596..3700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3768..3786
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tggcgaatcc tctgaccatt ccatttccac taatcttatc cctatcagtg atagagattg
       61 acatccctat cagtgataga gatactgagc acggatcccg actttaagaa ggagatatac
      121 atatggcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
      181 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
      241 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
      301 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
      361 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
      421 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
      481 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
      541 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
      601 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
      661 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
      721 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
      781 gtcactccac cggtgcttaa gccaggtaac gaatggatcc caaagcccgc cgaaaggcgg
      841 gcttttctgt tcttcacctt aatccattat acataactca tacattccaa actaatctaa
      901 cttcttccat cattaaccac acctctctat catcacctat tcccctctat ctccctcaaa
      961 ttctttcaca tattatatcc taacactcat acacccatac acaaatgtcg actctcaagg
     1021 acagaaaagc ccgcctttcg gcgggctttg agttcattag atcccggcgg cggtcacgaa
     1081 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     1141 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1201 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttggc
     1261 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctga tgtagttgta
     1321 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1381 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1441 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1501 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgcccc aggtcagggt
     1561 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     1621 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1681 tacgtcgccg tccagctcga ccaggatggg aacaacgcca gtgaaaagct ccattagaaa
     1741 gactcctaag catgattaag atgtttcagt acgaaaattg ctttcattgt tgatctcctt
     1801 tttaagtgaa cttgggcccg cttgctgtgc tcagtatctt gttatccgct cacaattgaa
     1861 cagcaatgct atgttgtggg atgaggtgga ttagtgggga taacgcagga aagaacatgt
     1921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2401 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     3361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3721 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3781 cgaggccctt tcgtcttcaa gaattc
//
