LOCUS       sequence-474305-        3704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M13 phagemid that expresses sgRNA-B1 from a strong constitutive 
            promoter. (This plasmid is a gift of the Jaramillo Lab, where it is 
            called PAJ615.).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3704)
  AUTHORS   Kushwaha M, Singh V, Jaramillo A
  TITLE     Development of a genetically-encoded in vivo computing platform to 
            play the game of Tic-Tac-Toe
  JOURNAL   Zenodo (2017) Conference: SB7.0: The Seventh International Meeting 
            on Synthetic Biology.
REFERENCE   2  (bases 1 to 3704)
  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: "Zenodo 
            (2017) Conference: SB7.0: The Seventh International Meeting on 
            Synthetic Biology."
COMMENT     Sequence Label: pSEVA63_sgRNA-B1_M13ps
FEATURES             Location/Qualifiers
     source          1..3704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..50
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        81..115
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      276..370
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(401..406,407..423,424..429)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             618..1151
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            1362..1470
                     /note="incP origin of transfer"
     CDS             complement(1481..2143)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2144..2915
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     rep_origin      complement(3019..3528)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3214..3235)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3422..3441
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(join(3631..3704,1..13))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttcgttttat ttgatgcctt taattaaagc ggataacaat ttcacacagg aggccgccta
       61 ggccgcggcc gcgcgaattc ttgacagcta gctcagtcct aggtataatg ctagctttac
      121 accccgcgtt tataggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cttttctgca ggcatgcaag cttgcggccg
      241 cgtcgtgact gggaaaaccc tggcgactag tcttggactc ctgttgatag atccagtaat
      301 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
      361 tggtgagaat ccaggggtcc ccaataatta cgatttaaat ttgacataag cctgttcggt
      421 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
      481 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
      541 acagcctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
      601 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
      661 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
      721 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
      781 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
      841 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
      901 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
      961 gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     1021 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     1081 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     1141 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     1201 aattggacaa cggtccgcgc gttgtccttt tccgctgcat aaccctgctt cggggtcatt
     1261 atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg
     1321 gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta
     1381 ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc
     1441 aacgggaatc ctgctctgcg aggctggccg taggccggcc ctaccggcgc ggcagcgtta
     1501 cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca tcgggcatcg
     1561 gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg
     1621 gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg gtcggtagtt
     1681 gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt
     1741 cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg cgcaactggt
     1801 cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg gtgccggggc
     1861 cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact gcgtattgga
     1921 ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc accacggcgt
     1981 tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg
     2041 caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct
     2101 tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc tccatgcggt
     2161 aggggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac
     2221 aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat
     2281 gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga
     2341 tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc
     2401 cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc
     2461 ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg
     2521 aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac
     2581 tgcccccact cgcataggct tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg
     2641 tcgctgcgcg agccttgacc cgccttccac ttggtgtcca accggcaagc gaagcgcgca
     2701 ggccgcaggc cggaggcttt tccccagaga aaattaaaaa aattgatggg gcaaggccgc
     2761 aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc
     2821 cctgtggtca agctcgtggg caggcgcagc ctgtccatca gcttgtccag cagggttgtc
     2881 cacgggccga gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc acatatccac
     2941 gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg
     3001 ggggcgcgag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     3061 gtataagcaa atatttaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3121 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3181 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3241 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3301 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3361 aaccctaaag ggagcccccg atttagagct tgacggggaa agcgaacgtg gcgagaaagg
     3421 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3481 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa aaggatctag
     3541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3601 tgaggcgcgc ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
     3661 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcct
//
