LOCUS       Exported                4440 bp ds-DNA     circular SYN 02-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   Barre Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 2, 2020 from SnapGene Viewer 5.1.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4440
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    46..302
                     /label=RP4 ori
     oriT            191..300
                     /note="incP origin of transfer"
     CDS             505..1551
                     /codon_start=1
                     /gene="tnp"
                     
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     misc_feature    1625..1651
                     /label=Himar-MmeI
     primer_bind     1630..1650
                     /label=3459
     CDS             2014..2808
                     /codon_start=1
                     /gene="kan"
                     /label=kan
                     /note="aminoglycoside 3'-phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     2793..2820
                     /label=1514
     misc_feature    2830..2863
                     /label=Rd2SP
     primer_bind     2847..2867
                     /label=3460
     protein_bind    2870..2903
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    2924..2957
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    complement(2961..2987)
                     /label=Himar-MmeI
     primer_bind     complement(2962..2982)
                     /label=3459
     rep_origin      3057..3445
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3570..4229)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4230..4332)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene"
ORIGIN
        1 caattgatat cgcgcgcgta atacgactca ctatagggcg aattgggtac cagcgctttt
       61 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
      121 tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac
      181 ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca
      241 ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg
      301 cgtcgacggt atcgataagc ttgatatcga attcccgcca gtgtgatgga ttgacacata
      361 gatggcgtcg ctagtattaa atgcatatta tttttatata gtaccaacct tcaaatgatt
      421 cgtgtcaaaa tttgacgtct gtaagtcaat tagtttgtga gatagagcgt cttttgtgaa
      481 gcaacttttg ttattgtgaa acatatggaa aaaaaggaat ttcgtgtttt gataaaatac
      541 tgttttctga agggaaaaaa tacagtggaa gcaaaaactt ggcttgataa tgagtttccg
      601 gactctgccc cagggaaatc aacaataatt gattggtatg caaaattcaa gcgtggtgaa
      661 atgagcacgg aggacggtga acgcagtgga cgcccgaaag aggtggttac cgacgaaaac
      721 atcaaaaaaa tccacaaaat gattttgaat gaccgtaaaa tgaagttgat cgagatagca
      781 gaggccttaa agatatcaaa ggaacgtgtt ggtcatatca ttcatcaata tttggatatg
      841 cggaagctct gtgcgaaatg ggtgccgcgc gagctcacat ttgaccaaaa acaacgacgt
      901 gttgatgatt ctaagcggtg tttgcagctg ttaactcgta atacacccga gtttttccgt
      961 cgatatgtga caatggatga aacatggctc catcactaca ctcctgagtc caatcgacag
     1021 tcggctgagt ggacagcgac cggtgaaccg tctccgaagc gtggaaagac tcaaaagtcc
     1081 gctggcaaag taatggcctc tgttttttgg gatgcgcatg gaataatttt tatcgattat
     1141 cttgagaagg gaaaaaccat caacagtgac tattatatgg cgttattgga gcgtttgaag
     1201 gtcgaaatcg cggcaaaacg gccccacatg aagaagaaaa aagtgttgtt ccaccaagac
     1261 aacgcaccgt gccacaagtc attgagaacg atggcaaaaa ttcatgaatt gggcttcgaa
     1321 ttgcttcccc acccaccgta ttctccagat ctggccccca gcgacttttt cttgttctca
     1381 gacctcaaaa ggatgctcgc agggaaaaaa tttggctgca atgaagaggt gatcgccgaa
     1441 actgaggcct attttgaggc aaaaccgaag gagtactacc aaaatggtat caaaaaattg
     1501 gaaggtcgtt ataatcgttg tatcgctctt gaagggaact atgttgaata ataaaaacga
     1561 attttgacaa aaaaatgtgt ttttctttgt tagaccggaa tcctccaatt cgggatccac
     1621 tagtacaggt tggatgataa gtccccggtc tttccgctag cttcacgctg ccgcaagcac
     1681 tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg
     1741 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
     1801 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
     1861 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     1921 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     1981 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2041 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2101 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2161 aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg
     2221 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2281 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2341 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2401 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2461 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2521 ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg atctcgtcgt gacccatggc
     2581 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     2641 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2701 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2761 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     2821 ggtacgcgtg tgactggagt tcagacgtgt gctcttccga tcttctagaa taacttcgta
     2881 tagcatacat tatacgaagt tatgaattcg gtacccggga ataataactt cgtatagcat
     2941 acattatacg aagttattct agaccgggga cttatcatcc aacctgtgcg gccgccaccg
     3001 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgcaa ttcccatgtc
     3061 agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag
     3121 tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt
     3181 atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt
     3241 atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     3301 ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt
     3361 agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac
     3421 aggttgaact gctgatcttc agatcctcta cgccggacgc atcgtggccg gatcagatct
     3481 gatatcgtcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     3541 tctaggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac
     3601 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     3661 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     3721 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     3781 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     3841 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     3901 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     3961 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga
     4021 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     4081 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     4141 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     4201 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     4261 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     4321 acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca
     4381 gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg
//
