LOCUS       sequence-250544-        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: pEYmRD3A
FEATURES             Location/Qualifiers
     source          1..3384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    270..286
                     /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    293..309
                     /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    662..680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    687..705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             746..1423
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1531..1549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1617..1721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1722..1790,1791..2582)
                     /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(1940..1959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2753..3341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3242..3261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgttc tttcctgcgt tatcctgtgg ttttgtaagg gggtaacaga
       61 aaagcccgcc tttcggcggg ctttgtggcg cccgaacagg gacttgaacc ctggacccgc
      121 ggccggacgc gactagttac ggagctcaca ctctactcaa caagctgcac tgccgaagca
      181 gccacacctg gacccgtcct tcaccatttc attcagttgc gtccggccgc tgctctaccg
      241 actgagctat ccgggctgtg ctcagtatct tgttatccgc tcacaatgtc aattgttatc
      301 cgctcacaat tgtatgggtg tatgagtgtt aggataaggt tggtgatgat aagatatttg
      361 ggtaatagaa gtgaaagatt agggtattta agagttattt agtattttaa ggaggaaagg
      421 aggaagagta agtttttaat atgtgaaaga atttgaggga gatagagggg aataggtgat
      481 gatagagagg tgtggttaat gatggaagaa gttagattag tttggaatgt atgagttagg
      541 ttggatttga tgggaagtat tgagatgggg tttaagtgta gggtaataat gatatataat
      601 tagattgatg ttaagaggaa tgggtaggta ttgagttatg tataatggat taaggtgaag
      661 atccctatca gtgatagaga ttgacatccc tatcagtgat agagatactg agcacggatc
      721 ccgactttaa gaaggagata tacatatggc gagtagcgaa gacgttatca aagagttcat
      781 gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga
      841 aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg
      901 tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta
      961 cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa
     1021 atgggaacgt gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc
     1081 cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga
     1141 cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc
     1201 ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca
     1261 ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc
     1321 ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga
     1381 acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taagccaggt aacgaatgga
     1441 tcccaaagcc cgccgaaagg cgggcttttc tgtcatccct aaacacacta attggtcaga
     1501 ggattcgcca gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1561 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     1981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2881 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3061 ggtctggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3361 ttttacggtt cctggccttt tgct
//
