LOCUS       sequence-224378-        7777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses scarlet tagged Kif5a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7777)
  TITLE     Koller Lab plasmids
REFERENCE   2  (bases 1 to 7777)
  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"
COMMENT     Sequence Label: pScarlet-Kif5a
FEATURES             Location/Qualifiers
     source          1..7777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      243..831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     732..751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1006..1309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1310..1513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1464..1484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1552..1561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1558..2253
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1691..1709)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1959..1978
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5510..5631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5547..5566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5601..5620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5638..6093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5775..5796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5987..6006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6120..6224
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6226..6583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6226..6417
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6249..6269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6434..6569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6618..7412
                     /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             6618..7412
                     /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(6672..6691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7282..7301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7600..7619)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7644..7691
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
       61 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
      121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      361 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      841 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      901 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
      961 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1021 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1081 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1141 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1201 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1261 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1321 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1381 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1441 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1501 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
     1561 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc
     1621 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1681 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc
     1741 ctgtcccctc agttcatgta cggctccagg gccttcacca agcaccccgc cgacatcccc
     1801 gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1861 gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac
     1921 aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca
     1981 atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac
     2041 attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc
     2101 tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac
     2161 atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc
     2221 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctcg actgaccatg
     2281 gcggagacta acaacgaatg cagcatcaag gtgctttgcc gatttcggcc cctgaaccag
     2341 gccgagattc tgcgggggga caagttcatc cccattttcc aaggggacga cagcgtcatt
     2401 attgggggaa agccatatgt ctttgaccgc gtcttccccc caaacaccac tcaggagcag
     2461 gtttaccacg cctgtgccat gcagatcgtc aaagacgtcc ttgctggtta caatggcaca
     2521 atcttcgctt atggacagac atcctcaggg aaaacgcata ccatggaggg gaagctgcac
     2581 gaccctcagc tgatgggcat cattccccgg atcgctcgag acatcttcaa ccacatctac
     2641 tccatggatg agaaccttga attccacatt aaggtatctt acttcgagat ttacctggat
     2701 aagatccgtg accttttgga tgtgaccaag acgaacctgt ccgtgcatga ggacaaaaac
     2761 cgggtgccgt ttgtcaaggg ttgtaccgaa cgctttgtgt ccagcccaga ggagattctg
     2821 gatgtgatcg atgaggggaa gtccaaccgt cacgtagctg tcaccaacat gaacgagcac
     2881 agttctcgga gccacagcat cttcctcatc aacatcaagc aggagaacgt agagaccgag
     2941 cagaagctca gcgggaagct gtacctcgtg gatctggccg gaagcgagaa ggtcagcaag
     3001 acaggggcag agggagccgt tctggacgag gcaaagaata tcaacaagtc gctgtcggcc
     3061 ctggggaacg tgatctctgc actggcagag ggcaccaaaa gctacgtgcc gtaccgcgac
     3121 agcaaaatga cgaggattct ccaggactct ctgggaggga actgcaggac taccatgttc
     3181 atctgctgct cgccgtccag ctacaatgac gcagagacca agtccacgct catgtttgga
     3241 cagcgggcga agaccatcaa gaacactgcc tcagtgaatc tggagctgac tgctgagcag
     3301 tggaagaaga agtatgagaa ggagaaggag aagaccaagg cccagaagga gacaattgcg
     3361 aagctagagg ctgagcttag ccggtggcgc aatggagaga atgtgcctga gactgagcgc
     3421 ctggctggag aggactcagc tctgggagct gagctctgcg aggagacccc tgtgaatgac
     3481 aactcatcca ttgtggtacg catcgcacct gaggaaaggc agaaatatga ggaagagatc
     3541 cgccgtctct acaagcagct tgatgacaag gatgatgaga tcaaccagca gagccagctc
     3601 attgagaagc tgaagcagca gatgctggac caggaagagc tgctcgtgtc cactcgggga
     3661 gacaacgaga aggtccagcg ggagcttagc cacctgcagt ccgagaacga tgctgcgaag
     3721 gacgaggtga aggaagtgct gcaggcccta gaggagctgg cggtcaacta cgaccagaag
     3781 tcccaggagg tggaggagaa gagccagcag aaccagctgc tggtggacga gctgtcccag
     3841 aaagtggcca ccatgctgtc cctggagtcc gagctacagc ggctccagga ggtcagtgga
     3901 caccagcgaa agcggatcgc tgaggtgctg aatgggctga tgagggacct gagtgagttc
     3961 agtgtcatcg tgggcaacgg cgagattaag ctgcccgtgg agatcagtgg ggccatcgag
     4021 gaggagttca cggtggcccg gctctacatc agcaagatca agtcggaggt gaagtccgtg
     4081 gttaagcgat gtcggcagct ggagaacctc caggtggagt gtcatcgcaa gatggaggtg
     4141 accggtaggg agctgtcatc ttgccaactg ctcatctcac agcatgaggc caagatccgt
     4201 tcactcacgg agtacatgca gactgtggag ttgaagaaac ggcacctgga agagtcctac
     4261 gactccctga gcgatgagct tgccaggctc caggcgcacg aaactgtaca cgaggtagct
     4321 ctgaaagaca aggagccaga cacacaggac gcggaggagg tgaagaaggc cctggaacta
     4381 cagatggaga atcaccgtga ggcccatcac cggcagctgg cccgcctccg ggatgagatt
     4441 aatgagaaac agaaaaccat tgatgagctg aaagacctga accagaagct ccagttagag
     4501 ctggagaagc ttcaggccga ctatgagagg ctgaagaatg aagagaacga gaagagcgcc
     4561 aagctccagg agctgacatt tctgtatgag cgacatgagc agtccaagca ggacctcaag
     4621 gggctggagg agacagttgc ccgtgaactc cagaccctcc acaaccttcg caagctgttc
     4681 gttcaagacg tcacgactcg agtcaagaaa agtgcagaaa tggagcccga ggacagtggg
     4741 gggattcatt cccaaaagca gaagatctcc tttcttgaga acaacctgga acagcttaca
     4801 aaggttcaca aacagctggt acgtgacaat gcagatctgc gttgtgagct tcctaaattg
     4861 gaaaaacgac ttcgggctac ggctgagaga gttaaggccc tggagggtgc actgaaggag
     4921 gccaaggagg gcgctatgaa ggacaagcgt agataccagc aggaggtgga ccgcatcaaa
     4981 gaagccgtgc ggtacaagag ctccggcaag cggggccatt ctgcccagat cgctaagcct
     5041 gtgaggcctg gccactatcc tgcctcctca cccaccaacc cctacggcac ccggagcccc
     5101 gagtgtatca gctacaccaa caacctcttc cagaactacc agaacctgca cctgcaggct
     5161 gcgcctagct ccacttcaga tatgtacttt gccagcagcg gagccacatc tgttgccccc
     5221 ttggcttcct accagaaggc caacatggac aatggaaatg ccacagatat caacgacaac
     5281 aggagtgacc tgccgtgtgg ctatgaggct gaggaccagg ccaagctttt ccctctccac
     5341 caagagacag cagccagcta actcccaacc ctggccacat ggatccaccg gatctagata
     5401 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5461 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5521 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5581 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     5641 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5701 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5761 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5821 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5881 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5941 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6001 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6061 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     6121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6181 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     6241 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     6301 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     6361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     6421 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6481 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6541 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     6601 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6781 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     7021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     7081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     7141 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7381 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7441 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7501 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7561 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     7621 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     7681 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     7741 gagaccccat tggggccaat acgcccgcgt ttcttcc
//
