LOCUS       sequence-393089-        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CJ8421_04975 Rptr2 targeted DNA sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  AUTHORS   Jiao C, Reckstadt C, Konig F, Homberger C, Yu J, Vogel J, Westermann
            AJ, Sharma CM, Beisel CL
  TITLE     RNA recording in single bacterial cells using reprogrammed 
            tracrRNAs.
  JOURNAL   Nat Biotechnol. 2023 Jan 5. doi: 10.1038/s41587-022-01604-8.
  PUBMED    36604543
REFERENCE   2  (bases 1 to 4894)
  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"; journalName: "Nat 
            Biotechnol. 2023 Jan 5. doi: 10.1038/s41587-022-01604-8."
COMMENT     Sequence Label: CJ8421_04975 Rptr2 recording plasmid
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(542..1492)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1540..1762
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2254..2348)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2379..3173)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2490..2509)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3100..3119
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3352..3370
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             3738..3760
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3797..3818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4058..4077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4405..4426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4711..4731
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      join(4812..4894,1..4)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaatccgccg ccctagacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt
       61 gttgctagtt tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg
      121 ctatttcttc cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt
      181 tgtcggcagc tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg
      241 ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact
      301 ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc
      361 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
      421 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
      481 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
      541 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
      601 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
      661 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
      721 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
      781 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
      841 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
      901 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
      961 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     1021 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     1081 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     1141 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     1201 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     1261 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     1321 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     1381 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     1441 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     1501 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     1561 tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     1621 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     1681 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     1741 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     1801 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     1861 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     1921 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     1981 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     2041 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     2101 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     2161 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     2221 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     2281 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2341 caggagtcca agcgagctct cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     2401 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     2461 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     2521 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     2581 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     2641 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     2701 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     2761 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     2821 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     2881 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     2941 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     3001 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     3061 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     3121 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     3181 gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc
     3241 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     3301 ggcaattccg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3361 atcacgaggc cctttcgtct tcaagaattc tggcgaatcc tctgaccagc cagaaaacga
     3421 cctttctgtg gtgaaaccgg atgctgcaat tcagagcgcc agcaagtggg ggacagcaga
     3481 agacctgacc gccgcagagt ggatgtttga catggtgaag actatcgcac catcagccag
     3541 aaaaccgaat tttgctgggt gggctaacga tatccgcctg atgcgtgaac gtgacggacg
     3601 taaccaccgc gacatgtgtg tgctgttccg ctgggcatgc taagttcaat gtttttacat
     3661 tgagaaacac tgagctaaca ccgtgcgtgt tgacaatttt acctctggcg gtgataatgg
     3721 ttgcagctag caataatttt gtttaacttt aagaaggaga tataccatgg agcttttcac
     3781 tggcgttgtt cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4441 ctaactcgag caaagcccgc cgaaaggcgg gcttttctgt gtcgaccgat gcccttgaga
     4501 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactaaact tcaaaattag
     4561 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     4621 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     4681 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     4741 gattacacat ggcatggatg aactatacaa ataaatgtcc agacctgcag gcatgcaagc
     4801 tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4861 gttgtttgtc ggtgaacgct ctcctgagta ggac
//
