LOCUS       sequence-258384-        2496 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 2496)
  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 2496)
  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: pEYC1A
FEATURES             Location/Qualifiers
     source          1..2496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    40..58
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    65..83
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    527..543
                     /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    550..566
                     /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(667..1255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(747..766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1426..2217,2218..2286))
                     /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     2049..2068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2287..2391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2459..2477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctggcgaatc ctctgaccat tccatttcca ctaatcttat ccctatcagt gatagagatt
       61 gacatcccta tcagtgatag agatactgag cacaggatcc cgactggcga gagccaggta
      121 acgaatggat cccaaagccc gccgaaaggc gggcttttct gttcttcacc ttaatccatt
      181 atacataact catacattcc aaactaatct aacttcttcc atcattaacc acacctctct
      241 atcatcacct attcccctct atctccctca aattctttca catattatat cctaacactc
      301 atacacccat acacagaaaa gcccgccttt cggcgggctt tgtggcgccc gaacagggac
      361 ttgaaccctg gacccgcggc cggacgcgac tagttacgga gctcacactc tactcaacaa
      421 gctgcactgc cgaagcagcc acacctggac ccgtccttca ccatttcatt cagttgcgtc
      481 cggccgctgc tctaccgact gagctatccg ggctgtgctc agtatcttgt tatccgctca
      541 caatgtcaat tgttatccgc tcacaattgg gatgaggtgg attagtgggg ataacgcagg
      601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1081 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2041 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2401 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2461 taggcgtatc acgaggccct ttcgtcttca agaatt
//
