LOCUS       sequence-273036-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Microtubules/Kinesin, C1 Cloning Vector, 
            Excitation:399, Emission:456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6911)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 6911)
  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: mTagBFP2-MCAK-C-7
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     383..402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        657..960
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        961..1164
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1115..1135
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1203..1212
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1209..1919
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    4119..4171
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4295..4416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4386..4405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4423..4878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4560..4581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4772..4791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4905..5009
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5011..5368
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5011..5202
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5034..5054)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5219..5354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5403..6197
                     /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             5403..6197
                     /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(5457..5476)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6067..6086
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6385..6404)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6429..6476
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6805..6911,1..482)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
       61 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
      601 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      661 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      721 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      781 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      841 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      901 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      961 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1021 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1081 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1141 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     1201 tcgccaccat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
     1261 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
     1321 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     1381 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
     1441 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
     1501 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     1561 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
     1621 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
     1681 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg
     1741 ccaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
     1801 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
     1861 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatt
     1921 ccggagccgc ttccgcctcc atggccatgg actcgtcgct tcaggcccgc ctgtttcccg
     1981 gtctcgctat caagatccaa cgcagtaatg gtttaattca cagtgccaat gtaaggactg
     2041 tgaacttgga gaaatcctgt gtttcagtgg aatgggcaga aggaggtgcc acaaagggca
     2101 aagagattga ttttgatgat gtggctgcaa taaacccaga actcttacag cttcttccct
     2161 tacatccgaa ggacaatctg cccttgcagg aaaatgtaac aatccagaaa caaaaacgga
     2221 gatccgtcaa ctccaaaatt cctgctccaa aagaaagtct tcgaagccgc tccactcgca
     2281 tgtccactgt ctcagagctt cgcatcacgg ctcaggagaa tgacatggag gtggagctgc
     2341 ctgcagctgc aaactcccgc aagcagtttt cagttcctcc tgcccccact aggccttcct
     2401 gccctgcagt ggctgaaata ccattgagga tggtcagcga ggagatggaa gagcaagtcc
     2461 attccatccg aggcagctct tctgcaaacc ctgtgaactc agttcggagg aaatcatgtc
     2521 ttgtgaagga agtggaaaaa atgaagaaca agcgagaaga gaagaaggcc cagaactctg
     2581 aaatgagaat gaagagagct caggagtatg acagtagttt tccaaactgg gaatttgccc
     2641 gaatgattaa agaatttcgg gctactttgg aatgtcatcc acttactatg actgatccta
     2701 tcgaagagca cagaatatgt gtctgtgtta ggaaacgccc actgaataag caagaattgg
     2761 ccaagaaaga aattgatgtg atttccattc ctagcaagtg tctcctcttg gtacatgaac
     2821 ccaagttgaa agtggactta acaaagtatc tggagaacca agcattctgc tttgactttg
     2881 catttgatga aacagcttcg aatgaagttg tctacaggtt cacagcaagg ccactggtac
     2941 agacaatctt tgaaggtgga aaagcaactt gttttgcata tggccagaca ggaagtggca
     3001 agacacatac tatgggcgga gacctctctg ggaaagccca gaatgcatcc aaagggatct
     3061 atgccatggc ctcccgggac gtcttcctcc tgaagaatca accctgctac cggaagttgg
     3121 gcctggaagt ctatgtgaca ttcttcgaga tctacaatgg gaagctgttt gacctgctca
     3181 acaagaaggc caagctgcgc gtgctggagg acggcaagca acaggtgcaa gtggtggggc
     3241 tgcaggagca tctggttaac tctgctgatg atgtcatcaa gatgatcgac atgggcagcg
     3301 cctgcagaac ctctgggcag acatttgcca actccaattc ctcccgctcc cacgcgtgct
     3361 tccaaattat tcttcgagct aaagggagaa tgcatggcaa gttctctttg gtagatctgg
     3421 cagggaatga gcgaggcgcg gacacttcca gtgctgaccg gcagacccgc atggagggcg
     3481 cagaaatcaa caagagtctc ttagccctga aggagtgcat cagggccctg ggacagaaca
     3541 aggctcacac cccgttccgt gagagcaagc tgacacaggt gctgagggac tccttcattg
     3601 gggagaactc taggacttgc atgattgcca cgatctcacc aggcataagc tcctgtgaat
     3661 atactttaaa caccctgaga tatgcagaca gggtcaagga gctgagcccc cacagtgggc
     3721 ccagtggaga gcagttgatt caaatggaaa cagaagagat ggaagcctgc tctaacgggg
     3781 cgctgattcc aggcaattta tccaaggaag aggaggaact gtcttcccag atgtccagct
     3841 ttaacgaagc catgactcag atcagggagc tggaggagaa ggctatggaa gagctcaagg
     3901 agatcataca gcaaggacca gactggcttg agctctctga gatgaccgag cagccagact
     3961 atgacctgga gacctttgtg aacaaagcgg aatctgctct ggcccagcaa gccaagcatt
     4021 tctcagccct gcgagatgtc atcaaggcct tgcgcctggc catgcagctg gaagagcagg
     4081 ctagcagaca aataagcagc aagaaacggc cccagtgact cgagctcaag cttcgaattc
     4141 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     4201 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4261 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4381 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     4441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4861 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4921 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4981 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     5041 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5101 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5161 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5221 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5281 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5341 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     5401 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5461 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5521 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5581 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5641 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5701 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5761 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5821 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5881 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5941 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6001 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6061 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6121 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6181 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6241 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6301 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6361 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6421 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6481 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6541 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6601 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6661 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6721 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6901 tctttttccg a
//
