LOCUS       40924_26669        3066 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKfloxANTSAsc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3066)
  AUTHORS   Yoon YG, Koob MD.
  TITLE     Nonreplicating Intracellular Bacterial Vector for Conjugative DNA 
            Transfer into Mitochondria
  JOURNAL   Pharm. Res. (2012) In press
  PUBMED    22350804
REFERENCE   2  (bases 1 to 3066)
  AUTHORS   Koob MD.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-DEC-2011) Lab Medicine and Pathology, ITN and IHG, 
            University of Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN 
            55455, USA
REFERENCE   3  (bases 1 to 3066)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3066)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Pharm. Res.
            (2012) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-DEC-2011) Lab Medicine and Pathology, ITN and IHG, University of
            Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN 55455, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..301
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(425..656)
                     /label=lambda attP
                     /note="integrase from phage lambda"
     rep_origin      complement(975..1563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1703..1736
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1757..2548)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      2790..2876
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2968..2995
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    complement(join(3013..3066,1..3))
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgctga tcagcgatga attaatggcg atgacgcatc
       61 ctcacgataa tatccgggta ggcgcaatca ctttcgtctc tactccgtta caaagcgagg
      121 ctgggtattt cccggccttt ctgttatccg aaatccactg aaagcacagc ggctggctga
      181 ggagataaat aataaacgag gggctgtatg cacaaagcat cttctgttga gttaagaacg
      241 agtatcgaga tggcacatag ccttgctcaa attggaatca ggtttgtgcc aataccagta
      301 gaaacagacg aagaagctag ctttgcactg gattgcgagg ctttgtgctt ctctggagtg
      361 cgacaggttt gatgacaaaa aattagcgca agaagacaaa aatcaccttg cgctaatgct
      421 ctgttacagg tcactaatac catctaagta gttgattcat agtgactgca tatgttgtgt
      481 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
      541 atcattttac gtttctcgtt cagctttttt atactaagtt ggcattataa aaaagcattg
      601 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgatt
      661 tcaattttgt cccactccct gcctctgtca tcacgatact gtgatgccat ggggcgcgcc
      721 agcggccgca ctagtcctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      781 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1381 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     1441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttttgcggc
     1681 cgcctagggg gccccccctc gaataacttc gtataatgta tgctatacga agttattcga
     1741 ggtcgaggcc tgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     1801 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     1861 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     1921 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     1981 cgccatgggt cacgacgaga tcctcgccgt cgggcatccg cgccttgagc ctggcgaaca
     2041 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     2101 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     2161 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     2221 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     2281 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     2341 gccacgatag ccgcgctgcc tcgtcttgga gttcattcag ggcaccggac aggtcggtct
     2401 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     2461 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     2521 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     2581 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     2641 agggcttccc aaccttacca gagggcgcac cagctggcaa ttcgtacgat aacttcgtat
     2701 aatgtatgct atacgaagtt attcgaatcg atggcccccc gatggtagtg tggggtctcc
     2761 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2821 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2881 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     2941 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     3001 ttggccagtg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag
     3061 ctcgaa
//
