LOCUS       sequence-97271-e        7367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a 2-integrase plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7367)
  AUTHORS   Yang L, Nielsen AA, Fernandez-Rodriguez J, McClune CJ, Laub MT, Lu 
            TK, Voigt CA
  TITLE     Permanent genetic memory with >1-byte capacity.
  JOURNAL   Nat Methods. 2014 Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Epub 
            2014 Oct 26.
  PUBMED    25344638
REFERENCE   2  (bases 1 to 7367)
  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: 
            "10.1038/nmeth.3147"; journalName: "Nat Methods"; date: "2014-12"; 
            volume: "11"; issue: "12"; pages: "1261-6"
COMMENT     Sequence Label: pCis_7+8
FEATURES             Location/Qualifiers
     source          1..7367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..84
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(580..599)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             591..1406
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             591..1406
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(664..683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1523..1540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      2114..2702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2603..2622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3184..3213)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(3260..4138)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4165..4449
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4372..4391
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 acttccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
       61 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      121 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      181 atttaacgcg aattttaaca aaatacttca ctgacaccct catcagtgcc aacatagtaa
      241 gccagtatac actccgctag cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat
      301 accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg ttgatgagag
      361 ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg gaacggtctg
      421 cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga tttattcaac
      481 aaagccacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
      541 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      601 ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat gctgatttat
      661 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
      721 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
      781 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
      841 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
      901 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
      961 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1021 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1081 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1141 agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1201 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1261 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1321 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1381 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     1441 gcattacgct gacttgacgg gactcgagat taccgccttt gagtgagctg ataccgctcg
     1501 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcctgca taacgcgaag
     1561 taatcttttc ggttttaaag aaaaagggca gggtggtgac accttgcccg tttttttgcc
     1621 ggactgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1681 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1741 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1801 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1861 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1921 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2041 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2221 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2821 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga ggtcggcgat ataggcgcca
     2881 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc
     2941 tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat
     3001 tctgcaaacc ctatgctact ccctcgagcc gtcaattgtc tgattcgtta ccaattatac
     3061 tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     3121 aagattactt cgcgtttgcc acctgacgtc taagaaaagg aatattcagc aatttgcccg
     3181 tgccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgctac gtaattagtt
     3241 agttagccct tagtgactct tatgacaact tgacggctac atcattcact ttttcttcac
     3301 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     3361 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     3421 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     3481 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     3541 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     3601 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     3661 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     3721 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     3781 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     3841 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     3901 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     3961 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     4021 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     4081 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     4141 ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc
     4201 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
     4261 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
     4321 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
     4381 catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg
     4441 tttctccata cccgtttttt tgggctagca gtaatttcaa caaaataact aggattcgaa
     4501 tgaaagtggc catttatgtt cgtgttagca ccgatgaaca ggccaaagaa ggttttagca
     4561 ttccggcaca gcgtgaacgt ctgcgtgcat tttgtgcaag ccagggttgg gaaattgtgc
     4621 aagaatatat tgaagaaggt tggagcgcaa aagatctgga tcgtccgcag atgcagcgtc
     4681 tgctgaaaga tatcaaaaaa ggcaacattg atattgtgct ggtgtatcgt ctggatcgcc
     4741 tgacccgtag cgttctggat ctgtatctgc tgctgcagac ctttgaaaaa tacaatgtgg
     4801 catttcgtag cgccaccgaa gtttatgata ccagcaccgc aatgggtcgt ctgtttatta
     4861 ccctggttgc agcactggca cagtgggaac gtgaaaatct ggcagaacgt gttaaatttg
     4921 gtatcgagca gatgatcgat gaaggtaaaa aaccgggtgg tcatagcccg tatggttaca
     4981 aatttgataa agacttcaat tgcaccatta ttgaggaaga agcagacgtt gttcgtatga
     5041 tctatcgcat gtattgtgat ggttatggct atcgtagcat tgcagatcgt ctgaatgaac
     5101 tgatggttaa accgcgtatt gccaaagaat ggaatcataa tagcgtgcgt gatatcctga
     5161 ccaacgatat ctatattggc acctatcgtt ggggtgataa agttgttccg aataatcatc
     5221 cgcctattat tagcgaaacc ctgttcaaaa aagcccagaa agaaaaagaa aaacgtggcg
     5281 ttgatcgtaa acgcgttggt aaatttctgt ttaccggtct gctgcagtgt ggtaattgtg
     5341 gtggccataa aatgcagggc cattttgata aacgtgagca gaaaacctat taccgttgta
     5401 ccaaatgtca ccgcattacc aacgaaaaaa acattctgga accgctgctg gatgaaattc
     5461 agctgctgat taccagcaaa gaatacttta tgagcaaatt cagcgaccgc tatgatcagc
     5521 aagaggttgt tgatgttagc gcactgacaa aagaactgga aaaaatcaaa cgccagaaag
     5581 agaaatggta cgatctgtat atggatgatc gtaacccgat tccgaaagaa gaactgtttg
     5641 ccaaaattaa cgaactgaac aaaaaagaag aagaaatcta tagcaagctg agcgaagtgg
     5701 aagaagataa agaaccggtt gaagagaaat ataaccgcct gagcaaaatg atcgatttta
     5761 aacagcagtt tgagcaggcc aacgacttta ccaaaaaaga gctgctgttc agcatcttcg
     5821 aaaagattgt gatttatcgc gagaaaggca agctgaaaaa aatcaccctg gattacaccc
     5881 tgaaataaga ataaaacaat aactaggatt cgaatgaaag ttgccgttta ttgtcgtgtt
     5941 agcaccctgg aacagaaaga acatggtcat agcattgaag aacaagagcg taaactgaaa
     6001 agcttctgcg atattaatga ttggaccgtg tatgatacct atatcgatgc aggttatagc
     6061 ggtgcaaaac gtgatcgtcc ggaactgcag cgtctgatga atgatattaa caaatttgat
     6121 ctggtgctgg tgtataaact ggatcgtctg acccgtaatg ttcgtgatct gctggacctg
     6181 ctggaaatct ttgaaaaaaa tgatgtgagc tttcgtagcg ccaccgaagt ttatgatacc
     6241 accaccgcaa tgggtcgtct gtttgttacc ctggttggtg caatggcaga atgggaacgt
     6301 gaaaccattc gtgaacgtac ccagatgggt aaactggcag cactgcgtaa aggtattatg
     6361 ctgaccaccc ctccgtttta ttatgaccgt gtggataata agtttgtgcc gaacaaatac
     6421 aaagacgtta ttctgtgggc atatgacgaa gcaatgaaag gtcagagcgc aaaagcaatt
     6481 gcacgcaaac tgaataatag cgatattccg cctccgaata atacccagtg gcagggtcgt
     6541 accattaccc atgccctgcg taatccgttt acccgtggtc attttgattg gggtggtgtg
     6601 catattgaaa ataaccatga accgatcatc accgatgaga tgtatgagaa agttaaagat
     6661 cgcctgaatg aacgcgtgaa caccaaaaaa gttcgtcata ccagcatttt tcgtggcaaa
     6721 ctggtttgtc cggtttgtaa tgcacgcctg accctgaata gccataaaaa gaaaagcaat
     6781 agcggctata tctttgtgaa acagtactac tgcaacaact gtaaagttac cccgaatctg
     6841 aaaccggtgt acatcaaaga aaaagaagtg attaaagttt tttacaatta tctgaaacgc
     6901 ttcgatctgg aaaaatatga ggttacccag aaacagaacg aaccggaaat caccatcgat
     6961 atcaataaag ttatggaaca gcgcaaacgc taccataaac tgtatgcaag cggtctgatg
     7021 caagaagatg aactgtttga cctgattaaa gaaaccgatc agaccattgc cgaatatgaa
     7081 aaacagaatg aaaaccgcga agtgaagcag tatgatatcg aagatatcaa acagtataaa
     7141 gatctgctgt tagaaatgtg ggatatcagc tccgatgaag ataaagagga ctttatcaaa
     7201 atggcgatta aaaacatcta ttttgaatat atcattggca ccggtaacac cagccgtaaa
     7261 cgtaatagcc tgaaaattac gagcattgaa ttctattaac agataaaaaa aatccttagc
     7321 tttcgctaag gatgatttct gattagggtg atggttcacg tagtggg
//
