LOCUS       sequence-267932-        4694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing synthetic overlapping gene, ilvA-acpP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Blazejewski T, Ho HI, Wang HH
  TITLE     Synthetic sequence entanglement augments stability and containment 
            of genetic information in cells.
  JOURNAL   Science. 2019 Aug 9;365(6453):595-598. doi: 10.1126/science.aav5477.
  PUBMED    31395784
REFERENCE   2  (bases 1 to 4694)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2019-08-9- 9"; volume: "365"; issue: 
            "6453"; pages: "595-598"
COMMENT     Sequence Label: pH9-ilvA-acp (DE2)
FEATURES             Location/Qualifiers
     source          1..4694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1584..1655
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1671..1698
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1726..1743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1860..2405)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1940..1959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2519..2613)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(2678..2772)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2803..3594)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVITEVSTQLSEVVGAIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(3962..4588)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     4493..4512
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        4604..4659
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4640..4658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4677..4688
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggctgact cgcaacccct gtccggtgct ccggaaggtg ccgaatattt aagagcagtg
       61 ctgcgcgcgc cggtttacga ggcggcgcag gttacgccgc tacaaaaaat ggaaaaactg
      121 tcgtcgcgtc ttgataacgt cattctggtg aagcgcgaag atcgccagcc agtgcacagc
      181 tttaagctgc gcggcgcata cgccatgatg gcgggcctga cggaagaaca gaaagcgcac
      241 ggcgtgatca ctgcttctgc gggtaaccac gcgcagggcg tcgcgttttc ttctgcgcgg
      301 ttaggcgtga aggccctgat cgttatgcca accgccaccg ccgacatcaa agtcgacgcg
      361 gtgcgcggct tcggcggcga agtgctgctc cacggcgcga actttgatga agcgaaagcc
      421 aaagcgatcg aactgtcaca gcagcagggg ttcacctggg tgccgccgtt cgaccatccg
      481 atggtgattg ccgggcaagg cacgctggcg ctggaactgc tccagcagga cgcccatctc
      541 gaccgcgtat ttgtgccagt cggcggcggc ggtctggctg ctggcgtggc ggtgctgatc
      601 aaacaactga tgccgcaaat caaagtgatc gccgtagaag cggaagactc cgcctgcctg
      661 aaagcagcgc tggatgcggg tcatccggtt gatctgccgc gcgtagggct atttgctgaa
      721 ggcgtagcgg taaaacgcat cggtgacgaa accttccgtt tatgccagga gtatctcgac
      781 gacatcatca ccgtcgatag cgatgcgatc tgtgcggcga tgaaggattt attcgaagat
      841 gtgcgcgcgg tggcggaacc ctctggcgcg ctggcgctgg cgggaatgaa aaaatatatc
      901 gccctgcaca acattcgcgg cgaacggctg gcgcatattc tttccggtgc caacgtgaac
      961 ttccacggcc tgcgctacgt ctcagaacgc tgcgaactgg gcgaacagcg tgaagcgttg
     1021 ttggctgtca ccatccctga agagaaagga tcattcctta agttttgcca attgttggga
     1081 gggaggagtg ttacggatgt caactataga agagcgggtg aagaagataa taggagagca
     1141 gttgggagtc aagcaagaag aagtgacgaa caacgcgagc ttcgtagaag acttaggagc
     1201 agacagttta gatacagtgg aattagtgat ggctctagag gaagagttcg atacagaaat
     1261 accggacgag gaggcagaaa agataacaac agttcaagcc gctatagact atataaacgg
     1321 acatcaagct tagtcccgga atcaccgggc gcgctgctgc gcttcctcaa cacgctgggt
     1381 acgtactgga acatttcttt gttccactat cgcagccatg gcaccgacta cgggcgcgta
     1441 ctggcggcgt tcgaacttgg cgaccatgaa ccggatttcg aaacccggct gaatgagctg
     1501 ggctacgatt gccacgacga aaccaataac ccggcgttca ggttcttttt ggcgggttag
     1561 ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1621 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     1681 cgggtgggcc tttctgcgtt tatacctagg gatatattcc gcttcctcgc tcactgactc
     1741 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     1801 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     1861 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     1981 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     2041 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     2101 cgaacccccc gttcagtccg acagctgcgc cttatccggt aactatcgtc ttgagtccaa
     2161 cccggaaaga catgcaaaag cacaactggc agcagccact ggtaattgat ttagaggagt
     2221 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     2281 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     2341 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     2401 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     2461 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat
     2521 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     2581 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
     2641 ccgccctgcc actcatcgca gtactactag tgcttggatt ctcaccaata aaaaacgccc
     2701 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     2761 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gcttatttgc cgactacctt
     2821 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     2881 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     2941 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     3001 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     3061 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     3121 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     3181 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     3241 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     3301 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     3361 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     3421 aagccttacg gtcaccgtaa ctagcaaatc aatatcactg tgtggcttca ggccgccatc
     3481 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatggc
     3541 gccaactacc tctgatagtt gagtcgatac ttcggtgatc accgcttccc tcataatgtt
     3601 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
     3661 acatcgaccc atggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc
     3721 aaaaaaacag tcatagcaag ccatgaaaac cgccactgca ccgttaccac cgctgcgttc
     3781 ggtcaaggtt ctggaccagt tgcgtgagcg catgcgaaac gatcctcatc ctgtctcttg
     3841 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     3901 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcgacgt
     3961 cttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc
     4021 cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata
     4081 atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt
     4141 gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat
     4201 tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact
     4261 gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag
     4321 cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag
     4381 ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc
     4441 gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt
     4501 tacgggttgt taaaccttcg attcctacct cattaagcag ctctaatgcg ctgttaatca
     4561 ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca ctctatcgtt
     4621 gatagagtta ttttaccact ccctatcagt gatagagaaa agaattcaaa agatctaaag
     4681 aggagaaagg atct
//
