LOCUS       sequence-198330-        4248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Copy number inducible plasmid. Contains mini-F origin for 
            maintenance and P1 lytic replication machinery under CopA promoter 
            for copy increase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4248)
  AUTHORS   Sheth RU, Yim SS, Wu FL, Wang HH
  TITLE     Multiplex recording of cellular events over time on CRISPR 
            biological tape.
  JOURNAL   Science. 2017 Dec 15;358(6369):1457-1461. doi: 
            10.1126/science.aao0958. Epub 2017 Nov 23.
  PUBMED    29170279
REFERENCE   2  (bases 1 to 4248)
  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.aao0958"; journalName: "Science"; date: 
            "2017-12-15- 15"; volume: "358"; issue: "6369"; pages: "1457-1461"
COMMENT     Sequence Label: pTrig-CopA
FEATURES             Location/Qualifiers
     source          1..4248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        459..508
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             539..550
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1690..1709)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1701..2516
                     /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             1701..2516
                     /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(1774..1793)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2604..2622
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2626..2669)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2674..2691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    2933..3183
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3186..3941)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      join(4032..4248,1..3)
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
ORIGIN
        1 aagggaagtt ttttcaggca tcgtgtgtaa gcagaatata taagtgctgt tccctggtgc
       61 ttcctcgctc actcgaaatt cccggggata gctttatgct tgtaaaccgt tttgtgaaaa
      121 aatttttaaa ataaaaaagg ggacctctag ggtccccaat taattagtaa tataatctat
      181 taaaggtcat tcaaaaggtc atccaccgga tcaattcccc tgctcgcgca ggctgggtgc
      241 caagctctcg ggtaacatca aggcccgatc cttggagccc ttgccctccc gcacgatgat
      301 cgtgccgtga tcgaaatcca gatccttgac ccgcagttgc aaaccctcac tgatccggac
      361 ttttacccgc ctggtttatt aatttcttga ccttcccctt gctggaaggt ttaaccttta
      421 tcacagccag tcaaaactgt cttaaaggag tgttttagct gtcaccggat gtgctttccg
      481 gtctgatgag tccgtgagga cgaaacagcc tctacaaata attttgttta atctagagaa
      541 agaggagaaa tactagatgc tggctaaagt cactttcctg agctgtataa cgatgagcga
      601 ttttactttt tctggctatg aattggcctg ctttgtaaca cactccggtc tatcccgtag
      661 cgccgggcat atcctgtcgc aatgtgcaaa tctcgcggca acaaccagtg aatacttcat
      721 tcacaagcct caccgcctga tcgcggcaga aactggttat agccaatcaa ccgtcgttcg
      781 tgcattccgt gaagctgtaa acaaaggtat cctgtctgta gagattgtta tcggcgatca
      841 ccgtgaacgt cgcgctaacc tgtaccggtt tacaccatcc tttttggcct tcgcacaaca
      901 agccaaaaat gcgctgatag aaagcaaatt aaagatctct tcagcggcaa ccaaggttaa
      961 agctgttctc gctaagacat tggctttatt taatttttta tccacacccc catgtcaaaa
     1021 tgataccccc tccccctgtc aggatgacgt ggcaataaag aataagaagt cacaagttaa
     1081 aaaaacaaaa agatcagttt ccggcggtgc cggaacaacc agcctcaaaa aattgacttc
     1141 atggatcgct aaggcaaaag caaaggctga caatctgcgg ttatccaaaa aacgcactca
     1201 aaaacatgag ttcaagcaga aagtagaggc ggctgcgcgg aaatatgctt acctgaagaa
     1261 caagcgttcg cctgatattg gcgggatatc aaacttcgat aacctaccgc attgcatgac
     1321 ggtaaacgaa gctcttaatg cggttttagc caaaaataaa gataacgaac aatggggtat
     1381 accggcagga ttcagagggt aatgaattgc tctaattata accatgcata ctttcaacac
     1441 ctctagtttg ccatgaggca aactcatagg tgtcctggta agaggacact gttgccaaaa
     1501 ctggacgccc cattattgca attaataaac aactaacgga caattctacc taacaataag
     1561 tggagttgcg gccggccgcg atccgctagc aaagccacgt tgtgtctcaa aatctctgat
     1621 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1681 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgaggccgc
     1741 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1801 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     1861 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1921 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1981 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     2041 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2101 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2161 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2221 ctgttgaaca agtctggaaa gaaatgcata aactgttgcc attctcaccg gattcagtcg
     2281 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2341 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2401 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2461 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2521 aattggttaa ttggttgtaa cactggcgag ctctgccacc tgacgtctaa gaaaccatta
     2581 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
     2641 aatccttagc tttcgctaag gatgatttct ggcctcgctc actgactcgc tgcgctcggt
     2701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatggcgc gccacgcgta
     2761 cccataatac ccataatagc tgtttgccat cgcgtatgca tcgatcacgt gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactcg gatcgatccg gcgcgcaccg
     2881 tgggaaaaac tccaggtaga ggtacacacg cggatagcca attcagagta ataaactgtg
     2941 ataatcaacc ctcatcaatg atgacgaact aacccccgat atcaggtcac atgacgaagg
     3001 gaaagagaag gaaatcaact gtgacaaact gccctcaaat ttggcttcct taaaaattac
     3061 agttcaaaaa gtatgagaaa atccatgcag gctgaaggaa acagcaaaac tgtgacaaat
     3121 taccctcagt aggtcagaac aaatgtgacg aaccaccctc aaatctgtga cagataaccc
     3181 tcagactatc ctgtcgtcat ggaagtgata tcgcggaagg aaaatacgat atgagtcgtc
     3241 tggcggcctt tctttttctc aatgtatgag aggcgcattg gagttctgct gttgatctca
     3301 ttaacacaga cttgcaggaa gcggcggcgg aagtcaggca tacgctggta actttgaggc
     3361 agctggtaac gctctatgat ccagtcgatt ttcagagaga cgatgcctga gccatccggc
     3421 ttacgatact gacacaggga ttcgtataaa cgcatggcat acggattggt gatttctttt
     3481 gtttcactaa gccgaaactg cgtaaaccgg ttctgtaacc cgataaagaa gggaatgaga
     3541 tatgggttga tatgtacact gtaaagccct ctggatggac tgtgcgcacg tttgataaac
     3601 caaggaaaag attcatagcc tttttcatcg ccggcatcct cttcagggcg ataaaaaacc
     3661 acttccttcc ccgcgaaact cttcaatgcc tgccgtatat ccttactggc ttccgcagag
     3721 gtcaatccga atatttcagc atatttagca acatggatct cgcagatacc gtcatgttcc
     3781 tgtagggtgc catcagattt tctgatctgg tcaacgaaca gatacagcat acgtttttga
     3841 tcccgggaga gactatatgc cgcctcagtg aggtcgtttg actggacgat tcgcgggcta
     3901 tttttacgtt tcttgtgatt gataaccgct gtttccgcca tgacagatcc atgtgaagtg
     3961 tgacaagttt ttagattgtc acactaaata aaaaagagtc aataagcagg gataactttg
     4021 tgaaaaaaca gcttcttctg agggcaattt gtcacagggt taagggcaat ttgtcacaga
     4081 caggactgtc atttgagggt gatttgtcac actgaaaggg caatttgtca caacaccttc
     4141 tgtagaacca gcatggataa aggcctacaa ggcgctctaa aaaagaagat ctaaaaacta
     4201 taaaaaaaat aattataaaa atatccccgt ggataagtgg ataacccc
//
