LOCUS       40924_41360        3860 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSTn5-KM, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3860)
  AUTHORS   Madhaiyan M, Peng N, Ngoh ST, Cheng H, Cai L, Fu L, Reddy C, Hong Y,
            Ji L.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 3860)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3860)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-APR-2013) Biomaterials "
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(25..58)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      59..143
                     /note="derived from mdh promoter in Methylobacterium 
                     radiotolerans"
                     /regulatory_class="promoter"
     promoter        147..191
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     CDS             214..1005
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     RBS             1016..1024
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      1043..1090
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    complement(1091..1124)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1210..1226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1234..1250)
                     /label=lac operator
                     /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)."
     promoter        complement(1258..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1303..1324)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2374..3231)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3232..3336)
                     /label=AmpR promoter
     primer_bind     3810..3826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3841..3859
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 tcgagggcct ttttggccac tagttaccgt tcgtatagca tacattatac gaagttatac
       61 ttttagaatt accttatcgt cccgacaaga aatgtccgac cagctaaaga cttctcctcc
      121 aatcaaagcc tagaaattat aggaaatcat aaaaaattta tttgctttgt gagcggataa
      181 caattataat aaacttgaaa aggaggtaaa catatgatcg agcaggacgg cctccacgcc
      241 ggctcgccgg ccgcgtgggt cgagcgcctc ttcggctacg actgggccca gcagaccatc
      301 ggctgctcgg acgccgccgt cttccgcctc tcggcgcagg gccgcccggt cctcttcgtc
      361 aagaccgacc tctcgggcgc cctcaacgag ctgcaggacg aggccgcccg cctctcgtgg
      421 ctcgccacca ccggcgtccc gtgcgccgcc gtcctcgacg tcgtcaccga ggccggccgc
      481 gactggctcc tcctcggcga ggtcccgggt caggacctcc tctcgtcgca cctcgccccg
      541 gccgagaagg tctccatcat ggccgacgcc atgcgccgcc tccacaccct cgacccggcc
      601 acctgcccgt tcgaccacca ggccaagcac cgcatcgagc gcgcccgcac ccgcatggag
      661 gccggcctcg tcgaccagga cgacctcgac gaggagcacc agggcctcgc cccggccgag
      721 ctcttcgccc gcctcaaggc ccgcatgccg gacggcgagg acctcgtcgt cacccacggc
      781 gacgcctgcc tcccgaacat catggtcgag aacggccgct tctcgggctt catcgactgc
      841 ggccgcctcg gcgtcgccga ccgctaccag gacatcgccc tcgccacccg cgacatcgcc
      901 gaggagctcg gcggcgagtg ggccgaccgc ttcctcgtcc tctacggcat cgccgccccg
      961 gactcgcagc gcatcgcctt ctaccgcctc ctcgacgagt tcttctaaaa atcattaagg
     1021 aggtaaaacc atggtttggt acctagcata accccttggg gcctctaaac gggtcttgag
     1081 gggttttttg ataacttcgt atagcataca ttatacgaac ggtagatatc aagctttcta
     1141 gaggatccct gcagttaatt aaagatctgt cgactgcaga ggcctgcatg caagcttggc
     1201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3361 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3421 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     3481 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3541 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     3601 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3661 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3721 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     3781 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgtttaaac
     3841 ctgtctctta tacacatctc
//
