LOCUS       sequence-207889-        2234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sortase in Ecoli off a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2234)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 2234)
  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: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1 sortase
FEATURES             Location/Qualifiers
     source          1..2234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     617..636
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        617..635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             667..689
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1147..1164
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1219..1237)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1233..1280
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1298..2113)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2021..2040
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2105..2124
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
      661 aataattttg tttaacttta agaaggagat atacatatgc aagctaaacc tcaaattccg
      721 aaagataaat caaaagtggc aggctatatt gaaattccag atgctgatat taaagaacca
      781 gtatatccag gaccagcaac acgcgaacaa ttaaatagag gtgtaagctt tgcagaagaa
      841 aatgaatcac tagatgatca aaatatttca attgcaggac acactttcat tgaccgtccg
      901 aactatcaat ttacaaatct taaagcagcc aaaaaaggta gtatggtgta ctttaaagtt
      961 ggtaatgaaa cacgtaagta taaaatgaca agtataagaa acgttaagcc aacagctgta
     1021 gaagttctag atgaacaaaa aggtaaagat aaacaattaa cattaattac ttgtgatgat
     1081 tacaatgaag agacaggcgt ttgggaaaca cgtaaaatct ttgtagctac agaagtcaaa
     1141 ctcgagcacc accaccacca ccactgagtc gaccggctgc taacaaagcc cgaaaggaag
     1201 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1261 gggtcttgag gggttttttg ctgaaagcca attctgatta gaaaaactca tcgagcatca
     1321 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1381 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1441 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1501 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1561 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1621 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     1681 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     1741 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1801 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     1861 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     1921 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     1981 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     2041 catataaatc agcatccatg ttggaattta atcgcggctt cgagcaagac gtttcccgtt
     2101 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     2161 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtgag
     2221 atcaaaggat cttc
//
