LOCUS       sequence-269506-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing cysJ-infA functional variant 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  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 4937)
  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: CI-4
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             74..85
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1918..1989
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2005..2032
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2060..2077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2194..2739)
                     /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(2274..2293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2853..2947)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(3012..3106)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3137..3928)
                     /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(4296..4922)
                     /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     4827..4846
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gttgacactc tatcgttgat agagtcattt taccactccc tatcagtgat agagaaaaga
       61 attcaaaaga tctaaagagg agaaaggatc tatgacgaca caggtcccac cttccgcgtt
      121 gcttccgttg aacccggagc aactggcacg ccttcaggcg gccacgaccg atttaactcc
      181 cacccagctt gcctgggttt ctggctattt ctggggcgta ctcaatcagc agcctgctgc
      241 gcttgcagcg acgccagcgc cagccgcaga aatgccgggt ataactatta tctccgcctc
      301 gcaaaccggc aatgcgcgcc gggttgctga agcattacgt gatgatttat tagcagcaaa
      361 actgaacgtt aagctggtga acgcgggcga ctataaattc aaacaaatcg ccagcgaaaa
      421 actgctcatc gtagtgacgt caacgcaagg ggaaggggaa ccgccggaag aagccgtcgc
      481 gctgcataag ttcctgttct ccaaaaaagc gccaaagctg gaaaacaccg cgtttgccgt
      541 gtttagcctc ggcgatagct cttatgaatt tttctgccag tccgggaaag atttcgacag
      601 caagctggcg gaactgggtg gtgaacgcct gctcgaccgt gtcgatgccg acgtagaata
      661 ccagaaagga ggatcagaat ggcgaaggaa gatagtagac gtgttgaagg cgaggttacc
      721 gagtgcctcc ccagcacaag ctgccgcgtc cgcctcagtg aacgaaaagt tgtcctcgtc
      781 accatacaca aaagaagctc ccttaacagc atctctatca gtgaatcaaa aaataacagg
      841 tcgggactca ctcaaagacg tccgtcacat agaaatatcg tatttagata gcgggatgcg
      901 ttaccagccg ggtgacgcgc tgggcgtctg gtatcagaac gatccggcac tggtgaaaga
      961 acttgtcgaa ctgctgtggc tgaaaggcga tgaacctgtc accgtcgagg gcaaaacgtt
     1021 gcctctgaac gaagcgctac agtggcactt cgaactgacc gtcaacaccg ccaacattgt
     1081 tgagaattac gccacgctta cccgcagtga aacactgctg ccgctggtgg gcgataaagc
     1141 gaagttacag cattacgccg cgacgacgcc gattgtcgac atggtgcgtt tctccccggc
     1201 acagcttgat gccgaagcgc taattaatct gctgcgcccg ctgacgccgc gtctgtattc
     1261 catcgcctcc tcgcaggcgg aagtcgagaa cgaagtacac gtcaccgttg gtgtggtgcg
     1321 ttacgacgtg gaaggccgcg cccgtgccgg tggtgcctcc agcttcctcg ctgaccgcgt
     1381 ggaagaagag ggcgaagtcc gcgtatttat cgaacataac gataacttcc gcctgccagc
     1441 caatccagaa accccggtga ttatgattgg cccaggcacc ggtattgcgc cgttccgcgc
     1501 ctttatgcag caacgcgccg ccgacgaagc gccaggtaaa aactggctgt tctttggtaa
     1561 tccgcacttt acggaagact tcctgtacca ggtggagtgg cagcgctacg tcaaagatgg
     1621 cgtgctgaca cgtatcgatc ttgcctggtc gcgcgatcaa aaagaaaaag tttacgtaca
     1681 agacaaactg cgcgaacagg gcgcggagct gtggcgctgg atcaatgatg gtgcccacat
     1741 ttatgtctgc ggcgacgcta atcgcatggc gaaagacgtt gagcaggcac ttctggaagt
     1801 gattgccgaa tttggtggca tggacaccga agcggcggat gaatttttaa gtgagctgcg
     1861 cgtagagcgc cgttatcagc gagatgtcta ctaactcgag taaggatctc caggcatcaa
     1921 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1981 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc
     2041 tagggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     2101 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     2161 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     2221 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     2281 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     2341 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     2401 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgacagct
     2461 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcacaac
     2521 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     2581 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     2641 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     2701 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     2761 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     2821 cccatacgat ataagttgtt actagtgctt ggattctcac caataaaaaa cgcccggcgg
     2881 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     2941 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtacta
     3001 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3061 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg
     3121 aaccccagag tcccgcttat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
     3181 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
     3241 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     3301 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     3361 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     3421 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     3481 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     3541 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     3601 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     3661 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     3721 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaactagca
     3781 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
     3841 ccagcaacgt cggttcgaga tggcgctcga tggcgccaac tacctctgat agttgagtcg
     3901 atacttcggt gatcaccgct tccctcataa tgtttaactt tgttttaggg cgactgccct
     3961 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccatggcg taacgcgctt
     4021 gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcatag caagccatga
     4081 aaaccgccac tgcaccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     4141 agcgcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc
     4201 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     4261 agggcgcccc agctggcaat tccgacgtcg acgtcttaag acccactttc acatttaagt
     4321 tgtttttcta atccgcatat gatcaattca aggccgaata agaaggctgg ctctgcacct
     4381 tggtgatcaa ataattcgat agcttgtcgt aataatggcg gcatactatc agtagtaggt
     4441 gtttcccttt cttctttagc gacttgatgc tcttgatctt ccaatacgca acctaaagta
     4501 aaatgcccca cagcgctgag tgcatataat gcattctcta gtgaaaaacc ttgttggcat
     4561 aaaaaggcta attgattttc gagagtttca tactgttttt ctgtaggccg tgtacctaaa
     4621 tgtacttttg ctccatcgcg atgacttagt aaagcacatc taaaactttt agcgttatta
     4681 cgtaaaaaat cttgccagct ttccccttct aaagggcaaa agtgagtatg gtgcctatct
     4741 aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat tttttacatg ccaatacaat
     4801 gtaggctgct ctacacctag cttctgggcg agtttacggg ttgttaaacc ttcgattccg
     4861 acctcattaa gcagctctaa tgcgctgtta atcactttac ttttatctaa tctagacatc
     4921 attaattcct aattttt
//
