LOCUS       62056_23235        2964 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2964)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..2964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(32..65)
                     /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)."
     CDS             126..938
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     protein_bind    complement(942..975)
                     /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)."
     CDS             complement(994..1011)
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     rep_origin      complement(1186..1574)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1679..2536)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2537..2641)
                     /label=AmpR promoter
     terminator      2732..2818
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2910..2937
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 agcttgtcga ccatcactag taaaactgca gtaccgttcg tatagcatac attatacgaa
       61 gttatggatt ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
      121 gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca
      181 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
      241 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
      301 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
      361 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
      421 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
      481 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
      541 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
      601 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
      661 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
      721 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
      781 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
      841 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
      901 ataaattgca gtttcatttg atgctcgatg agtttttcta aataacttcg tatagcatac
      961 attatacgaa cggtaggatc cgatatcagc catggaaccg cgtggcacca gggtacccag
     1021 atctgggctg tccatgtgct ggcgttcgaa tttagcagaa gcggtttctt tcatatggaa
     1081 ttccctctag agcggccgcc accgcggtgg agctcctgac gccgttggat acaccaagga
     1141 aagtctacac gggaggatat tcatatggac catggctaat tcccatgtca gccgttaagt
     1201 gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc
     1261 cctggcttgt tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt
     1321 tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt
     1381 attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag
     1441 agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa
     1501 catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg
     1561 ctgatcttca gatcctctac gccggacgca tcgtggccgg atcttgcggc cgcaaaaatt
     1621 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1681 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1741 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1801 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1861 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1921 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1981 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2041 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2101 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2161 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2221 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2281 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2341 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2401 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2461 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2521 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2581 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2641 gcacatttcc ccgaaaagtg ccacctgcat cgatggcccc ccgatggtag tgtggggtct
     2701 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2761 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2821 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2881 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2941 gtggccagtg ccaagcttgc atgc
//
