LOCUS       Exported                5861 bp ds-DNA     circular SYN 23-MAR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5861)
  AUTHORS   RWills
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5861
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..589
                     /label=CMV
                     /note="CMV"
     misc_feature    1
                     /label=GFP tag
                     /note="GFP tag"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     primer_bind     519..539
                     /label=CMV-F
     CDS             613..1320
                     /codon_start=1
                     /label=mCherry
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     613..630
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     616..633
                     /label=FPi-F2.0
     primer_bind     complement(616..633)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(663..681)
                     /label=mRFP-N1 (-)
     primer_bind     1175..1193
                     /label=AC41
     primer_bind     1253..1273
                     /label=mRFP-C1(+)
     primer_bind     1301..1320
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1301..1320)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     misc_feature    1330..1374
                     /label=MCS
                     /note="MCS"
     CDS             1375..2151
                     /codon_start=1
                     /label=Kinase
                     /translation="NKVSEGHVNFIIAYNMLTGIRVAVSRCSGIMKPLTPADFRFTKKL
                     AFDYHGNELTPSSQYAFKFKDYCPEVFRELRALFGLDPADYLVSLTSKYILSELNSPGK
                     SGSFFYYSRDYKYIIKTIHHSEHIHLRKHIQEYYNHVRDNPNTLICQFYGLHRVKMPIS
                     FQNKIKHRKIYFLVMNNLFPPHLDIHITYDLKGSTWGRFTNLDKERLAKDRSYRPVMKD
                     LNWLEEGQKIKFGPLKKKTFLTQLKKDVELLAKLNTM"
     primer_bind     complement(2127..2151)
                     /label=Mss4_D636_R
     primer_bind     2152..2184
                     /label=Mss4_D636K_F
     primer_bind     2154..2184
                     /label=Mss4_D636A_F
     CDS             2155..2514
                     /codon_start=1
                     /label=Kinase
                     /translation="YSLLIGIHDINKAKEDDLQLADTASIEEQPQTQGPIRTGTGTVVR
                     HFFREFEGGIRASDQFNNDVDLIYYVGIIDFLTNYSVMKKLETFWRSLRHDTKLVSAIP
                     PRDYANRFYEFIEDSV"
     misc_feature    2518..2556
                     /label=MCS
                     /note="MCS"
     primer_bind     complement(2555..2574)
                     /label=SV40-pArev
     misc_feature    2777..3232
                     /label=f1
                     /note="f1"
     promoter        3294..3322
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3406..3635
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3573..3708
                     /label=SV40
                     /note="SV40"
     misc_feature    3757..4551
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    4787..4805
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    5136..5779
                     /label=pUC
                     /note="pUC"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgaacaag
     1381 gtctcagagg gtcacgtgaa tttcatcatt gcctacaaca tgctcactgg cataagagtt
     1441 gccgtcagta gatgttctgg gatcatgaag ccactcaccc cagcagattt taggtttacc
     1501 aaaaaactgg cctttgatta tcacggcaat gagcttactc cgtcatccca gtatgcattt
     1561 aagttcaagg attattgtcc cgaggttttc cgggagctta gagcactgtt tggcctcgat
     1621 cctgccgatt atctggtgtc tctcacatct aaatacatcc tctcagaact gaatagccca
     1681 ggaaaatctg ggtcattttt ttattatagt cgcgattaca agtatattat taagactatt
     1741 caccactctg aacacatcca cctccggaaa cacatccaag aatactataa ccatgtccgc
     1801 gataacccga atactcttat ttgccaattt tatggcctgc atcgcgtcaa gatgccaatc
     1861 tccttccaga acaaaatcaa acatcgaaaa atatatttcc tggtgatgaa caacctgttc
     1921 cccccacatc ttgatattca catcacctat gatctcaagg gatccacatg gggcaggttc
     1981 accaatcttg acaaagagcg cttggctaaa gatcgcagct accgcccagt gatgaaagac
     2041 cttaactggc tggaggaggg tcaaaagatt aagttcgggc cactgaagaa aaagacattc
     2101 cttacccagc ttaagaagga tgtagagctt ctggccaaac tgaacacaat gaaatacagc
     2161 ctgcttattg gcatccacga tatcaataaa gccaaagagg acgacttgca attggcggac
     2221 actgctagca ttgaagaaca gcctcaaacg cagggcccta ttaggacagg cacaggcaca
     2281 gttgtacggc atttcttccg cgagttcgag ggcggcataa gagctagcga ccagtttaac
     2341 aacgacgtcg accttattta ttatgtgggg ataatcgact ttctgacaaa ctacagtgtc
     2401 atgaaaaaac tcgagacttt ttggcgctca cttcgacacg acactaaatt ggtcagcgct
     2461 atacctccac gggattatgc taatcgcttt tacgagttta tcgaggattc cgtctaacgg
     2521 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2581 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2641 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2761 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2821 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2881 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2941 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3001 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3061 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3121 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3181 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3301 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3361 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3721 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4261 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4561 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4741 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4801 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4861 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4921 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4981 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5281 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5821 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
