LOCUS       40924_26664        4436 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKfloxANTSAsc-tsOri-pBR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4436)
  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 4436)
  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 4436)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4436)
  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..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..301
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(406..1353)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(1401..1623)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     rep_origin      complement(2345..2933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3073..3106
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3127..3918)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      4160..4246
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4338..4365
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    complement(join(4383..4436,1..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 cccatggtaa cggtgaacag ttgttctact tttgtttgtt
      361 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
      421 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
      481 gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
      541 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga
      601 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
      661 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
      721 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
      781 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
      841 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
      901 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
      961 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     1021 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     1081 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     1141 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     1201 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     1261 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     1321 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     1381 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt
     1441 tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     1501 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     1561 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     1621 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     1681 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     1741 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     1801 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     1861 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     1921 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
     1981 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
     2041 agactggcta atgcacccag taaggcagcg gtatcatcaa caggcgcgcc agcggccgca
     2101 ctagtcctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     2161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2761 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttttgcggc cgcctagggg
     3061 gccccccctc gaataacttc gtataatgta tgctatacga agttattcga ggtcgaggcc
     3121 tgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     3181 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3241 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     3301 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     3361 cacgacgaga tcctcgccgt cgggcatccg cgccttgagc ctggcgaaca gttcggctgg
     3421 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3481 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3541 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3601 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     3661 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     3721 ccgcgctgcc tcgtcttgga gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     3781 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     3841 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     3901 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
     3961 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
     4021 aaccttacca gagggcgcac cagctggcaa ttcgtacgat aacttcgtat aatgtatgct
     4081 atacgaagtt attcgaatcg atggcccccc gatggtagtg tggggtctcc ccatgcgaga
     4141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4201 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     4261 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     4321 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttggccagtg
     4381 ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaa
//
