LOCUS       sequence-198329-        4301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Copy number inducible plasmid. Contains mini-F origin for 
            maintenance and P1 lytic replication machinery under Lac promoter 
            for copy increase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4301)
  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 4301)
  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
FEATURES             Location/Qualifiers
     source          1..4301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    423..444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(459..464,465..482,483..489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    497..513
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     502..524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     521..537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     521..537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1743..1762)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1754..2569
                     /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             1754..2569
                     /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(1827..1846)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2657..2675
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2679..2722)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2727..2744)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2881..2902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    2986..3236
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3239..3994)
                     /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(4085..4301,1..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 tgatccgatt
      361 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      421 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      481 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      541 gattacgcca agcgcggtac ctgtcgcggc aacgcgctaa cagacgtagt aagaaccacc
      601 agcattgtaa tgctggctaa agtcactttc ctgagctgta taacgatgag cgattttact
      661 ttttctggct atgaattggc ctgctttgta acacactccg gtctatcccg tagcgccggg
      721 catatcctgt cgcaatgtgc aaatctcgcg gcaacaacca gtgaatactt cattcacaag
      781 cctcaccgcc tgatcgcggc agaaactggt tatagccaat caaccgtcgt tcgtgcattc
      841 cgtgaagctg taaacaaagg tatcctgtct gtagagattg ttatcggcga tcaccgtgaa
      901 cgtcgcgcta acctgtaccg gtttacacca tcctttttgg ccttcgcaca acaagccaaa
      961 aatgcgctga tagaaagcaa attaaagatc tcttcagcgg caaccaaggt taaagctgtt
     1021 ctcgctaaga cattggcttt atttaatttt ttatccacac ccccatgtca aaatgatacc
     1081 ccctccccct gtcaggatga cgtggcaata aagaataaga agtcacaagt taaaaaaaca
     1141 aaaagatcag tttccggcgg tgccggaaca accagcctca aaaaattgac ttcatggatc
     1201 gctaaggcaa aagcaaaggc tgacaatctg cggttatcca aaaaacgcac tcaaaaacat
     1261 gagttcaagc agaaagtaga ggcggctgcg cggaaatatg cttacctgaa gaacaagcgt
     1321 tcgcctgata ttggcgggat atcaaacttc gataacctac cgcattgcat gacggtaaac
     1381 gaagctctta atgcggtttt agccaaaaat aaagataacg aacaatgggg tataccggca
     1441 ggattcagag ggtaatgaat tgctctaatt ataaccatgc atactttcaa cacctctagt
     1501 ttgccatgag gcaaactcat aggtgtcctg gtaagaggac actgttgcca aaactggacg
     1561 ccccattatt gcaattaata aacaactaac ggacaattct acctaacaat aagtggagtt
     1621 gcggccggcc gcgatccgct agcaaagcca cgttgtgtct caaaatctct gatgttacat
     1681 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1741 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgaggc cgcgattaaa
     1801 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1861 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1921 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1981 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2041 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     2101 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2161 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2221 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2281 acaagtctgg aaagaaatgc ataaactgtt gccattctca ccggattcag tcgtcactca
     2341 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2401 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2461 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2521 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2581 taattggttg taacactggc gagctctgcc acctgacgtc taagaaacca ttattatcat
     2641 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     2701 agctttcgct aaggatgatt tctggcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2761 ctgcggcgag cggtatcagc tcactcaaag gcggtaatgg cgcgccacgc gtacccataa
     2821 tacccataat agctgtttgc catcgcgtat gcatcgatca cgtgtccacg ttctttaata
     2881 gtggactctt gttccaaact ggaacaacac tcggatcgat ccggcgcgca ccgtgggaaa
     2941 aactccaggt agaggtacac acgcggatag ccaattcaga gtaataaact gtgataatca
     3001 accctcatca atgatgacga actaaccccc gatatcaggt cacatgacga agggaaagag
     3061 aaggaaatca actgtgacaa actgccctca aatttggctt ccttaaaaat tacagttcaa
     3121 aaagtatgag aaaatccatg caggctgaag gaaacagcaa aactgtgaca aattaccctc
     3181 agtaggtcag aacaaatgtg acgaaccacc ctcaaatctg tgacagataa ccctcagact
     3241 atcctgtcgt catggaagtg atatcgcgga aggaaaatac gatatgagtc gtctggcggc
     3301 ctttcttttt ctcaatgtat gagaggcgca ttggagttct gctgttgatc tcattaacac
     3361 agacttgcag gaagcggcgg cggaagtcag gcatacgctg gtaactttga ggcagctggt
     3421 aacgctctat gatccagtcg attttcagag agacgatgcc tgagccatcc ggcttacgat
     3481 actgacacag ggattcgtat aaacgcatgg catacggatt ggtgatttct tttgtttcac
     3541 taagccgaaa ctgcgtaaac cggttctgta acccgataaa gaagggaatg agatatgggt
     3601 tgatatgtac actgtaaagc cctctggatg gactgtgcgc acgtttgata aaccaaggaa
     3661 aagattcata gcctttttca tcgccggcat cctcttcagg gcgataaaaa accacttcct
     3721 tccccgcgaa actcttcaat gcctgccgta tatccttact ggcttccgca gaggtcaatc
     3781 cgaatatttc agcatattta gcaacatgga tctcgcagat accgtcatgt tcctgtaggg
     3841 tgccatcaga ttttctgatc tggtcaacga acagatacag catacgtttt tgatcccggg
     3901 agagactata tgccgcctca gtgaggtcgt ttgactggac gattcgcggg ctatttttac
     3961 gtttcttgtg attgataacc gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag
     4021 tttttagatt gtcacactaa ataaaaaaga gtcaataagc agggataact ttgtgaaaaa
     4081 acagcttctt ctgagggcaa tttgtcacag ggttaagggc aatttgtcac agacaggact
     4141 gtcatttgag ggtgatttgt cacactgaaa gggcaatttg tcacaacacc ttctgtagaa
     4201 ccagcatgga taaaggccta caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa
     4261 aataattata aaaatatccc cgtggataag tggataaccc c
//
