LOCUS       sequence-110810-        7775 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 7775)
  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 7775)
  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_2+5
FEATURES             Location/Qualifiers
     source          1..7775
                     /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 acatagcgaa tcgtaaggag ttaaaagatg
     4501 ccgattgcac cggaatttct gagcctggca tatccgggtc aagaatttcc ggcatatctg
     4561 tatggtcgtg caagccgtga tccgaaacgt aaaggtcgta gcgttcagag ccagctggat
     4621 gaaggtcgtg ccacctgtct ggatgcaggt tggccgattg ccggtgaatt taaagatgtt
     4681 gatcgtagcg caagcgcata tgcacgtcgt acccgtgatg aatttgaaga aatgattgca
     4741 ggtattcagg caggcgaatg tcgtattctg gttgcatttg aagcaagccg ttattatcgt
     4801 gatctggaag catatgttcg tctgcgtcgt gtttgtcgtg aagccggtgt tctgctgtgt
     4861 tataatggtc aggtttatga tctgagcaaa agcgcagatc gtaaagcaac cgcacaggat
     4921 gcagttaatg ccgaaggtga agcagatgat attcgtgaac gtaatctgcg taccacccgt
     4981 ctgaatgcaa aacgtggtgg tgcacatggt ccggttccgg atggttataa acgtcgttat
     5041 gatccggata gcggtgatct ggttgatcag attccgcatc ctgatcgtgc aggtctgatt
     5101 accgaaattt ttcgtcgtgc agcagcagcc gaaccgctgg cagcaatttg ccgtgatctg
     5161 aatgaacgtg gtgaaaccac ccatcgtggt aaagcatggc agcgtcatca tctgcatgca
     5221 attctgcgta atcctgcata tattggtcat cgtcgtcatc tgggtgttga taccggtaaa
     5281 ggtatgtggg caccgatttg tgatgatgaa gattttgcag aaacctttca ggccgttcaa
     5341 gaaattctga gtctgcctgg tcgtcagctg agtccgggtc cggaagcaca gcatctgcag
     5401 accggtattg cactgtgtgg tgaacatcct gatgaaccgc ctctgcgtag cgttaccgtt
     5461 cgtggtcgta ccaattataa ctgtagcacc cgttatgatg ttgccatgcg tgaagatcgt
     5521 atggatgcat ttgttgaaga aagcgtgatt acctggctgg caagtgatga agcagttgca
     5581 gcctttgaag ataacaccga tgatgaacgt acccgtaaag cacgtattcg tctgaaagtt
     5641 ctggaagaac agctggaagc agcacagaaa caggcacgta ccctgcgtcc ggatggcatg
     5701 ggtatgctgc tgagcattga tagcctggca ggtctggaag ccgaactgac accgcagatt
     5761 gataaagccc gtcaagaaag ccgtagcctg catgttccgg cactgctgcg tgatctgctg
     5821 ggtaaaccgc gtgcagatgt ggatcgtgca tggaatgaag cactgaccct gccgcagcgt
     5881 cgcatgattc tgcgcatggt tgttaccatt cgtctgttta aagcaggtag ccgtggtgtt
     5941 cgtgcaattg aaccgggtcg tattaccctg agctatgttg gcgaaccggg ttttaaaccg
     6001 gttggtggta atcgtgcaaa acagtaagaa tcagaggaag gaggctcgat gcctggtatg
     6061 accaccgaaa ccggtccgga tcctgcaggt ctgattgacc tgttttgtcg taaaagcaaa
     6121 gcagttaaaa gccgtgcaaa tggtgcaggt cagcgtcgta aacaagaaat tagcattgca
     6181 gcacaagaaa ccctgggtcg taaagttgca gcactgctgg gtatgcaggt tcgtcatgtt
     6241 tggaaagaag ttggtagcgc aagccgtttt cgtaaaggta aagcacgtga tgatcagagc
     6301 aaagcactga aagccctgga aagcggtgaa gttggtgcac tgtggtgtta tcgtctggat
     6361 cgttgggatc gtggtggtgc cggtgcaatt ctgaaaatta tcgaaccgga agatggtatg
     6421 cctcgtcgtc tgctgtttgg ttgggatgaa gataccggtc gtccggttct ggatagcacc
     6481 aataaacgtg atcgcggtga actgattcgt cgtgcagaag aagcacgcga agaagcagaa
     6541 aaactgagcg aacgtgttcg tgataccaaa gcacatcagc gtgaaaatgg tgaatgggtt
     6601 aatgcccgtg caccgtatgg tctgcgtgtt gttctggtta ccgttagtga tgaagagggt
     6661 gatgaatatg atgaacgtaa actggcagca gatgatgaag atgcgggtgg tcctgatggt
     6721 ctgaccaaag cagaagcagc ccgtctggtt tttaccctgc cggttaccga tcgtctgagc
     6781 tatgcaggca ccgcacatgc aatgaatacc cgtgaaattc cgagcccgac cggtggtccg
     6841 tggattgcag ttaccgtgcg tgatatgatt cagaatccgg catatgcggg ttggcagacc
     6901 acaggtcgtc aggatggtaa acagcgtcgt ctgacctttt ataacggtga aggtaaacgt
     6961 gttagcgtta tgcatggtcc tccgctggtg accgatgaag aacaagaagc cgcaaaagca
     7021 gccgttaaag gtgaagatgg tgttggtgtt ccgctggatg gtagcgatca tgatacccgt
     7081 cgcaaacatc tgctgagcgg tcgtatgcgt tgtccgggtt gtggtggtag ctgtagctat
     7141 agcggtaatg gttatcgttg ttggcgtagc agtgtgaaag gtggttgtcc ggcaccgacc
     7201 tatgttgcac gtaaaagcgt tgaagaatat gttgcatttc gttgggcagc aaaattagca
     7261 gcaagcgaac cggatgatcc gtttgttatt gcagttgcag atcgctgggc agcactgacc
     7321 catccgcagg caagcgaaga tgaaaagtat gcaaaagccg cagttcgtga agccgaaaaa
     7381 aatctgggtc gcctgctgcg tgatcgtcag aatggtgttt atgatggtcc ggcagaacag
     7441 ttttttgccc ctgcatatca agaagcactg agcaccctgc aggcagccaa agatgcagtt
     7501 agcgaaagca gcgcaagcgc agcagttgat gttagctgga ttgttgatag cagcgattat
     7561 gaagaactgt ggctgcgtgc aaccccgacc atgcgtaatg caattattga tacctgcatc
     7621 gatgaaattt gggttgcaaa aggccagcgt ggtcgtccgt ttgatggtga tgaacgcgtt
     7681 aaaatcaaat gggcagcccg tacctaacag ataaaaaaaa tccttagctt tcgctaagga
     7741 tgatttctga ttagggtgat ggttcacgta gtggg
//
