LOCUS       sequence-272270-        6144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expresion of tdTomato SKAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pDK714
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        147..450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        451..654
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     605..625
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1460..1469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1466..2896
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    3018..3139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3055..3074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3109..3128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3146..3601)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3283..3304)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3495..3514
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3628..3732
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3734..4091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3734..3925
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3757..3777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3942..4077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4126..4920
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4126..4920
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4180..4199)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4790..4809
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5108..5127)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5152..5199
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5528..6116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6017..6036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
       61 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
      121 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      181 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      241 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      301 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      361 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      421 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      481 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      541 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      601 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      661 gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag cttcgaattc
      721 tgcagtcgac gccaccatga ccagtgtggt taagacagtg tatagcctgc agcccccctc
      781 tgcgctgagc ggcggccagc cggcagacac acaaactcgg gccacttcta agagtctctt
      841 acctgttagg tccaaagaag tcgatgtttc caaacagctt cattcaggag gtccagagaa
      901 tgatgttaca aaaatcacca aactgagacg agagaatggg caaatgaaag ctactgacac
      961 tgccaccaga aggaatgtca gaaagggata taagccttta tccaagcaaa aatcagagga
     1021 agagctcaag gacaagaacc agctgttaga agccgtcaac aagcagttgc accagaagtt
     1081 gactgaaact cagggagagc tgaaggacct gacccagaag gtagagctgc tggagaagtt
     1141 tcgggacaac tgtttggcaa ttttggagag caagggcctt gatccagctt taggcagtga
     1201 gaccctggca tcacgacaag aatccactac tgatcacatg gactctatgt tgctgttaga
     1261 aactttgcaa gaggagctga agctttttaa cgaaacagcc aaaaagcaga tggaggagtt
     1321 acaggcctta aaggtaaagc tggagatgaa agaggagcgc gtgaggtttt tggaacagca
     1381 aaccttatgt aacaatcaag taaatgattt aacaacagcc cttaaggaaa tggagcagct
     1441 attagaaatg ggaccggtcg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt
     1501 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
     1561 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1621 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
     1681 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
     1741 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     1801 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     1861 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     1921 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     1981 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     2041 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     2101 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     2161 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa
     2221 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     2281 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2341 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     2401 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     2461 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     2521 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     2581 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     2641 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     2701 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     2761 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     2821 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga
     2881 cgagctgtac aagtaggcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2941 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3001 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3121 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3181 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3241 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3301 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3361 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3421 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3481 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3541 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3601 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3661 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3721 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4021 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4081 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4141 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4201 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4261 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4321 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4381 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4441 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4501 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4561 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4621 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4681 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4741 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4801 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4861 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4921 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4981 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5041 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5101 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5161 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5221 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5281 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5341 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5401 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5461 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6121 ccagcaacgc ggccttttta cggt
//
