LOCUS       sequence-269509-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing cysJ-infA functional variant 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  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 6275)
  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-6
FEATURES             Location/Qualifiers
     source          1..6275
                     /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(3532..4077)
                     /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(3612..3631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4191..4285)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(4350..4444)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4475..5266)
                     /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(5634..6260)
                     /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     6165..6184
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gttgacactc tatcgttgat agagttattt 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 ggcgaaagaa gatagtagaa gtgttgaagg caagagtacc
      721 agctgcctcc ccagcacaag ctgccgcgtc cgctgcagtg aacagcacga tccatacgtc
      781 tccctacaca aaagaagctc ccttaacagc atctctagca gtgaatcaaa aaataacagg
      841 tcgggactcg gacaaagacg tccgtcacat agaaatagcg 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 acactgatga atcccctaat gatttttatc aaaatcatta agttaaggta gatacacatc
     2221 ttgtcatatg atcaaatggt ttcgccaaaa atcaataatc agacaacaaa atgtgcgaac
     2281 tcgatatttt acacgactct ctttaccaat tctgccccga attacactta aaacgactca
     2341 acagcttaac gttggcttgc cacgccttac ttgactgtaa aactctcact cttaccgaac
     2401 ttggccgtaa cctgccaacc aaagcgagaa caaaacataa catcaaacga atcgaccgat
     2461 tgttaggtaa tcgtcacctc cacaaagagc gactcgctgt ataccgttgg catgctagct
     2521 ttatctgttc gggcaatacg atgcccattg tacttgttga ctggtctgat atccgtgagc
     2581 aaaaacggct tatggtattg cgagcttcag tcgcactaca cggtcgttct gttactcttt
     2641 atgagaaagc gttcccgctt tcagagcaat gttcaaagaa agctcatgac caatttctag
     2701 ccgaccttgc gagcattcta ccgagtaaca ccacaccgct cattgtcagt gatgctggct
     2761 ttaaagtgcc atggtataaa tccgttgaga agctgggttg gtactggtta agtcgagtaa
     2821 gaggaaaagt acaatatgca gacctaggag cggaaaactg gaaacctatc agcaacttac
     2881 atgatatgtc atctagtcac tcaaagactt taggctataa gaggctgact aaaagcaacc
     2941 caatctcatg ccaaattcta ttgtataaat ctcgctctaa aggccgaaaa aatcagcgct
     3001 cgacacggac tcattgtcac cacccgtcac ctaaaatcta ctcagcgtcg gcaaaggagc
     3061 catggattct agcaactaac ttacctgttg aaattcgaac acccaaacaa cttgttaata
     3121 tctattcgaa gcgaatgcag attgaagaaa ccttccgaga cttgaaaagt cctgcctacg
     3181 gactaggcct acgccatagc cgaacgagca gctcagagcg ttttgatatc atgctgctaa
     3241 tcgccctgat gcttcaacta acatgttggc ttgcgggcgt tcatgctcag aaacaaggtt
     3301 gggacaagca cttccaggct aacacagtca gaaatcgaaa cgtactctca acagttcgct
     3361 taggcatgga agttttgcgg cattctggct acacaataac aagggaagac tcactcgtgg
     3421 ctgcaaccct gcttactcaa aatctattca cacatggtta cgttttgggg aaattatgag
     3481 gggatctctc agtacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     3541 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     3601 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     3661 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     3721 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     3781 ccgttcagtc cgacagctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     3841 gacatgcaaa agcacaactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     3901 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     3961 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     4021 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     4081 gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
     4141 gtagcacctg aagtcagccc catacgatat aagttgttac tagtgcttgg attctcacca
     4201 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     4261 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     4321 ccactcatcg cagtactact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     4381 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     4441 agtccaagcg agctctcgaa ccccagagtc ccgcttattt gccgactacc ttggtgatct
     4501 cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
     4561 cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca
     4621 ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc
     4681 tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg
     4741 gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg
     4801 gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg
     4861 tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt
     4921 cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga
     4981 tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg
     5041 aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta
     5101 cggtcaccgt aactagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg
     5161 agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg gcgccaacta
     5221 cctctgatag ttgagtcgat acttcggtga tcaccgcttc cctcataatg tttaactttg
     5281 ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
     5341 ccatggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac
     5401 agtcatagca agccatgaaa accgccactg caccgttacc accgctgcgt tcggtcaagg
     5461 ttctggacca gttgcgtgag cgcatgcgaa acgatcctca tcctgtctct tgatcagatc
     5521 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     5581 gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtcgac gtcttaagac
     5641 ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag gccgaataag
     5701 aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc
     5761 atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc
     5821 aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt
     5881 gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct
     5941 gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta
     6001 aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag
     6061 tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt
     6121 tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt
     6181 gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt
     6241 ttatctaatc tagacatcat taattcctaa ttttt
//
