LOCUS       sequence-426213-        5973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Imaging of Tubulin using Photoactivatable GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  TITLE     Imaging of KIF5B and Tubulin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5973)
  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            "Unpublished"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: PA-GFP-alpha-Tubulin
FEATURES             Location/Qualifiers
     source          1..5973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1314..1435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1351..1370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1405..1424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1442..1897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1579..1600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1791..1810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1924..2028
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2053..2073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2097..2307
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2119..2315
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2166..2301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2350..3144
                     /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             2350..3144
                     /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(2404..2423)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3014..3033
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3332..3351)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3376..3423
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3752..4340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4241..4260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4515..4818
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4819..5022
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4973..4993
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5061..5070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5067..5069,5070..5072,5073..5783)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5112..5133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5373..5392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5720..5741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctggcaagca cgtgccccgg gctgtgtttg tagacttgga acccacagtc attgatgaag
       61 ttcgcactgg cacctaccgc cagctcttcc accctgagca gctcatcaca ggcaaggaag
      121 atgctgccaa taactatgcc cgagggcact acaccattgg caaggagatc attgaccttg
      181 tgttggaccg aattcgcaag ctggctgacc agtgcaccgg tcttcagggc ttcttggttt
      241 tccacagctt tggtggggga actggttctg ggttcacctc cctgctcatg gaacgtctct
      301 cagttgatta tggcaagaag tccaagctgg agttctccat ttacccagca ccccaggttt
      361 ccacagctgt agttgagccc tacaactcca tcctcaccac ccacaccacc ctggagcact
      421 ctgattgtgc cttcatggta gacaatgagg ccatctatga catctgtcgt agaaacctcg
      481 atatcgagcg cccaacctac actaacctta accgccttat tagccagatt gtgtcctcca
      541 tcactgcttc cctgagattt gatggagccc tgaatgttga cctgacagaa ttccagacca
      601 acctggtgcc ctacccccgc atccacttcc ctctggccac atatgcccct gtcatctctg
      661 ctgagaaagc ctaccatgaa cagctttctg tagcagagat caccaatgct tgctttgagc
      721 cagccaacca gatggtgaaa tgtgaccctc gccatggtaa atacatggct tgctgcctgt
      781 tgtaccgtgg tgacgtggtt cccaaagatg tcaatgctgc cattgccacc atcaaaacca
      841 agcgcagcat ccagtttgtg gattggtgcc ccactggctt caaggttggc atcaactacc
      901 agcctcccac tgtggtgcct ggtggagacc tggccaaggt acagagagct gtgtgcatgc
      961 tgagcaacac cacagccatt gctgaggcct gggctcgcct ggaccacaag tttgacctga
     1021 tgtatgccaa gcgtgccttt gttcactggt acgtgggtga ggggatggag gaaggcgagt
     1081 tttcagaggc ccgtgaagat atggctgccc ttgagaagga ttatgaggag gttggtgtgg
     1141 attctgttga aggagagggt gaggaagaag gagaggaata ctaaggatcc accggatcta
     1201 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1261 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1321 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1381 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     1441 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1501 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1561 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1621 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1681 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     1741 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     1801 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     1861 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     1921 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1981 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2041 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2101 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2161 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2221 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2281 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     2341 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2401 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2461 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2521 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2581 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     2641 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     2701 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     2761 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     2821 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     2881 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     2941 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3001 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3061 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3121 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3181 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3241 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3301 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3361 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     3421 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3481 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3541 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     3601 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3661 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3721 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3781 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3841 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3901 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3961 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4021 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4081 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4141 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4201 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4261 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4321 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4381 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4441 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4501 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4561 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4621 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4681 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4741 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4801 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4861 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     4921 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     4981 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     5041 atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     5101 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     5161 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     5221 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cagctacggc gtgcagtgct
     5281 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     5341 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     5401 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     5461 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     5521 atatcatggc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     5581 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     5641 gccccgtgct gctgcccgac aaccactacc tgagccacca gtccgccctg agcaaagacc
     5701 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     5761 tcggcatgga cgagctgtac aagtccggac tcagatctcg agtgcgtgag tgcatctcca
     5821 tccacgttgg ccaggctggt gtccagattg gcaatgcctg ctgggagctc tactgcctgg
     5881 aacacggcat ccagcccgat ggccagatgc caagtgacaa gaccattggg ggaggagatg
     5941 actccttcaa caccttcttc agtgagacgg gcg
//
