LOCUS       sequence-229075-        7777 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1248
                     /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(686..704)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     954..973
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4505..4626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4542..4561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4596..4615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4633..5088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4770..4791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4982..5001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5115..5219
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5221..5578
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5221..5412
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5244..5264)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5429..5564
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5613..6407
                     /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             5613..6407
                     /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(5667..5686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6277..6296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6595..6614)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6639..6686
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7015..7603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7504..7523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag
      601 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      661 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      721 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt caccaagcac
      781 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
      841 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
      901 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
      961 atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc
     1021 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg
     1081 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc
     1141 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa
     1201 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
     1261 tctcgactga ccatggcgga gactaacaac gaatgcagca tcaaggtgct ttgccgattt
     1321 cggcccctga accaggccga gattctgcgg ggggacaagt tcatccccat tttccaaggg
     1381 gacgacagcg tcattattgg gggaaagcca tatgtctttg accgcgtctt ccccccaaac
     1441 accactcagg agcaggttta ccacgcctgt gccatgcaga tcgtcaaaga cgtccttgct
     1501 ggttacaatg gcacaatctt cgcttatgga cagacatcct cagggaaaac gcataccatg
     1561 gaggggaagc tgcacgaccc tcagctgatg ggcatcattc cccggatcgc tcgagacatc
     1621 ttcaaccaca tctactccat ggatgagaac cttgaattcc acattaaggt atcttacttc
     1681 gagatttacc tggataagat ccgtgacctt ttggatgtga ccaagacgaa cctgtccgtg
     1741 catgaggaca aaaaccgggt gccgtttgtc aagggttgta ccgaacgctt tgtgtccagc
     1801 ccagaggaga ttctggatgt gatcgatgag gggaagtcca accgtcacgt agctgtcacc
     1861 aacatgaacg agcacagttc tcggagccac agcatcttcc tcatcaacat caagcaggag
     1921 aacgtagaga ccgagcagaa gctcagcggg aagctgtacc tcgtggatct ggccggaagc
     1981 gagaaggtca gcaagacagg ggcagaggga gccgttctgg acgaggcaaa gaatatcaac
     2041 aagtcgctgt cggccctggg gaacgtgatc tctgcactgg cagagggcac caaaagctac
     2101 gtgccgtacc gcgacagcaa aatgacgagg attctccagg actctctggg agggaactgc
     2161 aggactacca tgttcatctg ctgctcgccg tccagctaca atgacgcaga gaccaagtcc
     2221 acgctcatgt ttgggcagcg ggcgaagacc atcaagaaca ctgcctcagt gaatctggag
     2281 ctgactgctg agcagtggaa gaagaagtat gagaaggaga aggagaagac caagacccag
     2341 aaggagacaa ttgcgaagct agaggctgag cttagccggt ggcgcaatgg agagaatgtg
     2401 cctgagactg agcgcctggc tggagaggac tcagctctgg gagctgagct ctgcgaggag
     2461 acccctgtga atgacaactc atccattgtg gtacgcatcg cacctgagga aaggcagaaa
     2521 tatgaggaag agatccgccg tctctacaag cagcttgatg acaaggatga tgagatcaac
     2581 cagcagagcc agctcattga gaagctgaag cagcagatgc tggaccagga agagctgctc
     2641 gtgtccactc ggggagacaa cgagaaggtc cagcgggagc ttagccacct gcagtccgag
     2701 aacgatgctg cgaaggacga ggtgaaggaa gtgctgcagg ccctagagga gctggcggtc
     2761 aactacgacc agaagtccca ggaggtggag gagaagagcc agcagaacca gctgctggtg
     2821 gacgagctgt cccagaaagt ggccaccatg ctgtccctgg agtccgagct acagcggctc
     2881 caggaggtca gtggacacca gcgaaagcgg atcgctgagg tgctgaatgg gctgatgagg
     2941 gacctgagtg agttcagtgt catcgtgggc aacggcgaga ttaagctgcc cgtggagatc
     3001 agtggggcca tcgaggagga gttcacggtg gcccggctct acatcagcaa gatcaagtcg
     3061 gaggtgaagt ccgtggttaa gcgatgtcgg cagctggaga acctccaggt ggagtgtcat
     3121 cgcaagatgg aggtgaccgg tagggagctg tcatcttgcc aactgctcat ctcacagcat
     3181 gaggccaaga tccgttcact cacggagtac atgcagactg tggagttgaa gaaacggcac
     3241 ctggaagagt cctacgactc cctgagcgat gagcttgcca ggctccaggc gcacgaaact
     3301 gtacacgagg tagctctgaa agacaaggag ccagacacac aggacgcgga ggaggtgaag
     3361 aaggccctgg aactacagat ggagaatcac cgtgaggccc atcaccggca gctggcccgc
     3421 ctccgggatg agattaatga gaaacagaaa accattgatg agctgaaaga cctgaaccag
     3481 aagctccagt tagagctgga gaagcttcag gccgactatg agaggctgaa gaatgaagag
     3541 aatgagaaga gcgccaagct ccaggagctg acatttctgt atgagcgaca tgagcagtcc
     3601 aagcaggacc tcaaggggct ggaggagaca gttgcccgtg aactccagac cctccacaac
     3661 cttcgcaagc tgttcgttca agacgtcacg actcgagtca agaaaagtgc agaaatggag
     3721 cccgaggaca gtggggggat tcattcccaa aagcagaaga tctcctttct tgagaacaac
     3781 ctggaacagc ttacaaaggt tcacaaacag ctggtacgtg acaatgcaga tctgcgttgt
     3841 gagcttccta aattggaaaa acgacttcgg gctacggctg agagagttaa ggccctggag
     3901 ggtgcactga aggaggccaa ggagggtgct atgaaggaca agcgtagata ccagcaggag
     3961 gtggaccgca tcaaggaagc cgtgcggtac aagagctccg gcaagcgggg ccattctgcc
     4021 cagatcgcta agcctgtgag gcctggccac tatcctgcct cctcacccac caacccctac
     4081 ggcacccgga gccccgagtg tatcagctac accaacaacc tcttccagaa ctaccagaac
     4141 ctgcacctgc aggctgcgcc tagctccact tcagatatgt actttgccag tagcggagcc
     4201 acatctgttg cccccttggc ttcctaccag aaggccaaca tggacaatgg aaatgccaca
     4261 gatatcaacg acaacaggag tgacctgccg tgtggctatg aggctgagga ccaggccaag
     4321 cttttccctc tccaccaaga gacagcagcc agctaactcc caaccctggc cacatggatc
     4381 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4441 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4501 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4621 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4681 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4741 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4801 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4861 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4921 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4981 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5041 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5221 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5281 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5401 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5461 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5521 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5581 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5641 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5701 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5761 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5821 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5881 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5941 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6001 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6061 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6121 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6181 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6241 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6301 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6361 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6421 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6481 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6541 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6601 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6661 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6721 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6781 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6841 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6901 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6961 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7021 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7081 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7141 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7201 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7261 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7321 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7381 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7441 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7501 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7561 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7621 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7681 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     7741 cggggtcatt agttcatagc ccatatatgg agttccg
//
