LOCUS       sequence-258357-        3828 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 3828)
  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 3828)
  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-D1A
FEATURES             Location/Qualifiers
     source          1..3828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      487..1075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     976..995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1248..1269)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1597..1616
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1856..1877
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2012..2028
                     /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    2035..2051
                     /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    2224..2242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2249..2267
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2308..2985
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3093..3111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3179..3283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3284..3352,3353..3828,1..316)
                     /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     complement(3502..3521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
       61 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      121 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      181 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      241 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      301 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
      361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      601 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      781 ggataaggcg cagcggtctg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1081 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1141 tcctgcgtta tcctgtggtt ttgtaagggg gtaacaaatg tcgactctca aggacagaaa
     1201 agcccgcctt tcggcgggct ttgagttcat tagatcccgg cggcggtcac gaactccagc
     1261 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     1321 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     1381 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     1441 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgatgtagtt gtactccagc
     1501 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     1561 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     1621 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     1681 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cccaggtcag ggtggtcacg
     1741 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     1801 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     1861 ccgtccagct cgaccaggat gggaacaacg ccagtgaaaa gctccattag aaagactcct
     1921 aagcatgatt aagatgtttc agtacgaaaa ttgctttcat tgttgatctc ctttttaagt
     1981 gaacttgggc ccgcttgctg tgctcagtat cttgttatcc gctcacaatg tcaattgtta
     2041 tccgctcaca attgaacagc aatgctatgt tgtgtatggg tgtatgagtg ttaggatata
     2101 atatgtgaaa gaatttgagg gagatagagg ggaataggtg atgatagaga ggtgtggtta
     2161 atgatggaag aagttagatt agtttggaat gtatgagtta tgtataatgg attaaggtga
     2221 agatccctat cagtgataga gattgacatc cctatcagtg atagagatac tgagcacgga
     2281 tcccgacttt aagaaggaga tatacatatg gcgagtagcg aagacgttat caaagagttc
     2341 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
     2401 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
     2461 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
     2521 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
     2581 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
     2641 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
     2701 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
     2761 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
     2821 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
     2881 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     2941 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaagccag gtaacgaatg
     3001 gatcccaaag cccgccgaaa ggcgggcttt tctgtcatcc ctaaacacac taattggtca
     3061 gaggattcgc cagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3121 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3301 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3361 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3421 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3481 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3541 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3601 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3661 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3721 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3781 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgt
//
