LOCUS       sequence-424730-        6505 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Centrosomes, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6505)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 6505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mCherry-PLK1-N-16
FEATURES             Location/Qualifiers
     source          1..6505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             159..953
                     /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             159..953
                     /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(213..232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     823..842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1141..1160)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1185..1232
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1561..2149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2050..2069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2324..2627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2628..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2782..2802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4717..4726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4723..5433
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4868..4886)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5136..5155
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5556..5677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5593..5612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5647..5666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5684..6139)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5821..5842)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6033..6052
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6166..6270
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6272..6505,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6272..6463
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6295..6315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6480..6505,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      181 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      241 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      301 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      361 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      421 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      481 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      541 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      601 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      661 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
      721 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
      781 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
      841 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
      901 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
      961 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1021 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1081 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     1141 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     1201 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     1261 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     1321 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     1381 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     1441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1681 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1741 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1801 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1861 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1921 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1981 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2041 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2101 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2161 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2221 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     2281 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2341 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2401 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2461 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2521 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2581 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2641 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2701 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2761 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2821 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ccaccatgag tgctgcagtg
     2881 actgcaggga agctggcacg ggcaccggcc gaccctggga aagccggggt ccccggagtt
     2941 gcagctcccg gagctccggc ggcggctcca ccggcgaaag agatcccgga ggtcctagtg
     3001 gacccacgca gccggcggcg ctatgtgcgg ggccgctttt tgggcaaggg cggctttgcc
     3061 aagtgcttcg agatctcgga cgcggacacc aaggaggtgt tcgcgggcaa gattgtgcct
     3121 aagtctctgc tgctcaagcc gcaccagagg gagaagatgt ccatggaaat atccattcac
     3181 cgcagcctcg cccaccagca cgtcgtagga ttccacggct ttttcgagga caacgacttc
     3241 gtgttcgtgg tgttggagct ctgccgccgg aggtctctcc tggagctgca caagaggagg
     3301 aaagccctga ctgagcctga ggcccgatac tacctacggc aaattgtgct tggctgccag
     3361 tacctgcacc gaaaccgagt tattcatcga gacctcaagc tgggcaacct tttcctgaat
     3421 gaagatctgg aggtgaaaat aggggatttt ggactggcaa ccaaagtcga atatgacggg
     3481 gagaggaaga agaccctgtg tgggactcct aattacatag ctcccgaggt gctgagcaag
     3541 aaagggcaca gtttcgaggt ggatgtgtgg tccattgggt gtatcatgta taccttgtta
     3601 gtgggcaaac caccttttga gacttcttgc ctaaaagaga cctacctccg gatcaagaag
     3661 aatgaataca gtattcccaa gcacatcaac cccgtggccg cctccctcat ccagaagatg
     3721 cttcagacag atcccactgc ccgcccaacc attaacgagc tgcttaatga cgagttcttt
     3781 acttctggct atatccctgc ccgtctcccc atcacctgcc tgaccattcc accaaggttt
     3841 tcgattgctc ccagcagcct ggaccccagc aaccggaagc ccctcacagt cctcaataaa
     3901 ggcttggaga accccctgcc tgagcgtccc cgggaaaaag aagaaccagt ggttcgagag
     3961 acaggtgagg tggtcgactg ccacctcagt gacatgctgc agcagctgca cagtgtcaat
     4021 gcctccaagc cctcggagcg tgggctggtc aggcaagagg aggctgagga tcctgcctgc
     4081 atccccatct tctgggtcag caagtgggtg gactattcgg acaagtacgg ccttgggtat
     4141 cagctctgtg ataacagcgt gggggtgctc ttcaatgact caacacgcct catcctctac
     4201 aatgatggtg acagcctgca gtacatagag cgtgacggca ctgagtccta cctcaccgtg
     4261 agttcccatc ccaactcctt gatgaagaag atcaccctcc ttaaatattt ccgcaattac
     4321 atgagcgagc acttgctgaa ggcaggtgcc aacatcacgc cgcgcgaagg tgatgagctc
     4381 gcccggctgc cctacctacg gacctggttc cgcacccgca gcgccatcat cctgcacctc
     4441 agcaacggca gcgtgcagat caacttcttc caggatcaca ccaagctcat cttgtgccca
     4501 ctgatggcag ccgtgaccta catcgacgag aagcgggact tccgcacata ccgcctgagt
     4561 ctcctggagg agtacggctg ctgcaaggag ctggccagcc ggctccgcta cgcccgcact
     4621 atggtggaca agctgctgag ctcacgctcg gccagcaacc gtctcaaggc ctccggtagc
     4681 ggcagcgcat ccggcggaag cggaagcgga ccggtcgcca ccatggtgag caagggcgag
     4741 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     4801 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     4861 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     4921 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     4981 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     5041 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     5101 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     5161 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     5221 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     5281 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     5341 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     5401 tccaccggcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     5461 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     5521 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     5581 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5641 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     5701 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5761 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5821 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5881 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5941 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6001 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6061 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6121 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6241 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     6301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6481 tcccgcccct aactccgccc agttc
//
