LOCUS       pSpyB-Tn7sg_Bank        3292 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3292)
  AUTHORS   ISU Biology IT
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..63
                     /label=PS1
     misc_feature    56..142
                     /label=rrnB T1 terminator
     promoter        join(157..161,162..167,168..184,185..190,191..197)
                     /label=BG35 Promoter
     misc_feature    239..258
                     /label=input into CRISPOR
     misc_feature    239..258
                     /label=Tn7sg
                     /note="sgRNA targeting the Tn7-R region of the mini-Tn7 
                     transposon
                     "
     misc_RNA        259..334
                     /label=Spy gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      336..379
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      468..570
                     /label=lambda t0 terminator
     primer_bind     complement(526..542)
                     /label=PS2
     CDS             698..1513
                     /codon_start=1
                     /label=KanR
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    1680..1788
                     /label=oriT
     CDS             complement(1799..2461)
                     /codon_start=1
                     /label=pBBR1 rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2462..3233
                     /label=pBBR1 oriV
ORIGIN
        1 gaagcccgga gagcaagccc gtaggggggg cgcgcccagc tgtctagggc ggcggatttg
       61 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
      121 actgagcctt tcgttttatt tgatgccttt aattaattta tttgacatgc gtgatgttta
      181 gaattataat ttggggacct aggccgcggc cgcgcgaatt cgagctcggt ctcatcacgt
      241 atgctttttc acagcatagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      301 ttatcaactt gaaaaagtgg caccgagtcg gtgctataaa acgaaaggcc cagtctttcg
      361 actgagcctt tcgttttata actaggtacc cggggatcct ctagagtcga cctgcaggca
      421 tgcaagcttg cggccgcgtc gtgactggga aaaccctggc gactagtctt ggactcctgt
      481 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
      541 gccgggcgtt ttttattggt gagaatccag gggtccccaa taattacgat ttaaatttgt
      601 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
      661 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc agcgtgaaac
      721 gagctgtagc cgtccgcgtc tgaacagcaa catggatgcg gatctgtatg gctataaatg
      781 ggcgcgtgat aacgtgggtc agagcggcgc gaccatttat cgtctgtatg gcaaaccgga
      841 tgcgccggaa ctgtttctga aacatggcaa aggcagcgtg gcgaacgatg tgaccgatga
      901 aatggtgcgt ctgaactggc tgaccgaatt tatgccgctg ccgaccatta aacattttat
      961 tcgcaccccg gatgatgcgt ggctgctgac caccgcgatt ccgggcaaaa ccgcgtttca
     1021 ggtgctggaa gaatatccgg atagcggcga aaacattgtg gatgcgctgg ccgtgtttct
     1081 gcgtcgtctg catagcattc cggtgtgcaa ctgcccgttt aacagcgatc gtgtgtttcg
     1141 tctggcccag gcgcagagcc gtatgaacaa cggcctggtg gatgcgagcg attttgatga
     1201 tgaacgtaac ggctggccgg tggaacaggt gtggaaagaa atgcataaac tgctgccgtt
     1261 tagcccggat agcgtggtga cccacggcga ttttagcctg gataacctga ttttcgatga
     1321 aggcaaactg attggctgca ttgatgtggg ccgtgtgggc attgcggatc gttatcagga
     1381 tctggccatt ctgtggaact gcctgggcga atttagcccg agcctgcaaa aacgtctgtt
     1441 tcagaaatat ggcattgata atccggatat gaacaaactg caatttcatc tgatgctgga
     1501 tgaatttttc taataattaa ttggaccgcg gtccgcgcgt tgtccttttc cgctgcataa
     1561 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     1621 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     1681 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     1741 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggccct
     1801 accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac
     1861 acggctcatc gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg
     1921 tcaaggctgg caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc
     1981 cggggccggt cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc
     2041 catgccccaa cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca
     2101 ccgacaggcg caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg
     2161 ccgacacggt gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt
     2221 ccttgactgc gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct
     2281 ggctgaccac cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg
     2341 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
     2401 gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
     2461 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
     2521 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
     2581 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
     2641 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
     2701 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
     2761 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
     2821 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
     2881 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
     2941 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac
     3001 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
     3061 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
     3121 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
     3181 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
     3241 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gc
//
