LOCUS       sequence-368851-        8094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat myosin 1b isoform b labelled with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8094)
  AUTHORS   Raposo G, Cordonnier MN, Tenza D, Menichi B, Durrbach A, Louvard D, 
            Coudrier E
  TITLE     Association of myosin I alpha with endosomes and lysosomes in 
            mammalian cells.
  JOURNAL   Mol Biol Cell. 1999 May . 10(5):1477-94.
  PUBMED    10233157
REFERENCE   2  (bases 1 to 8094)
  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: "Mol Biol 
            Cell. 1999 May . 10(5):1477-94."
COMMENT     Sequence Label: EGFP-Myo1b
FEATURES             Location/Qualifiers
     source          1..8094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4882..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4973..4992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5010..5465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5359..5378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5492..5596
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5598..5955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5598..5789
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5621..5641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5806..5941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5990..6784
                     /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             5990..6784
                     /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(6044..6063)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6654..6673
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6972..6991)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7016..7063
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7392..7980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7881..7900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attcggatcc tagatatcca
     1381 cccatggcca agaaggaggt aaaatcctca cttctggaca atatgatcgg cgttggagat
     1441 acggttctct tggagcctct caacgaggag acgttcatcg acaacctgaa gaagcgcttc
     1501 gatcacaatg agatatatac atacatcggg agtgtggtta tatctgttaa cccctaccgg
     1561 tccttgccca tttattcacc agagaaagtg gaagattaca gaaatagaaa cttctatgaa
     1621 ctgagccctc acatctttgc tttgtcggat gaagcataca gatccctccg tgatcaagat
     1681 aaagatcagt gcattctcat tactggggag agtggagcgg gaaaaacaga ggccagcaag
     1741 cttgtgatgt cctatgtcgc tgcagtgtgt ggaaaaggag cagaagttaa tcaggtcaag
     1801 gaacagcttc tccagtccac ccctgtcctg gaagcttttg gaaatgccaa gactgtaagg
     1861 aatgacaatt catccagatt cggcaaatac atggatattg aatttgactt taaaggcgat
     1921 ccgctgggag gagtgataag taactatctt ttagagaaat ctcgtgttgt taagcagccg
     1981 agaggtgaaa gaaacttcca cgtgttctat cagctgctct cgggtgcctc tgaggaactc
     2041 ctccacaagc ttaaactgga gcgggatttc agcagataca actatctgag tctggactct
     2101 gccaaagtga atggtgtgga cgacgctgcc aatttcagaa ctgtcaggaa tgccatgcag
     2161 atcgtgggct tttcggatcc tgaggccgag tctgtcctgg aggttgtggc agctgtcttg
     2221 aaactgggga acatcgagtt caagcctgaa tcacgaatga acggtttgga tgaaagcaaa
     2281 atcaaagaca aaaatgagtt aaaagaaatt tgtgagttga ccagcatcga tcaagtggtt
     2341 ttagaaagag cgttcagttt ccgaacggtt gaggcgaagc aggagaaagt ttcaaccaca
     2401 ttgaatgtgg ctcaggctta ttatgcccgt gatgccctgg ctaaaaacct ctacagccgg
     2461 ttgttttcat ggttggtaaa tcgaatcaac gaaagcatta aggcacaaac caaagtgaga
     2521 aagaaggtca tgggtgttct ggacatttat ggctttgaga tatttgagga caacagcttc
     2581 gagcagttca ttattaatta ttgtaatgaa aagcttcaac aaatcttcat tgaactgacc
     2641 ctcaaagaag agcaagagga atatattcgt gaggacatag aatggactca cattgactac
     2701 ttcaacaatg ctatcatttg cgacctaata gaaaacaaca caaacggaat cttggccatg
     2761 ctggatgaag agtgcctgag acccggcact gttacagatg agaccttcct ggaaaagctg
     2821 aaccaagtct gtgccaccca ccagcacttc gagagcagga tgagcaaatg ttcccgcttc
     2881 ctcaatgaca cgaccctgcc tcacagctgc ttcagaatcc agcattatgc tggcaaggtg
     2941 ctataccagg tggaagggtt tgttgacaag aacaatgacc ttctgtaccg agacctgtct
     3001 caagccatgt ggaaggctgg ccatgccctc atcaagtctc tgttccctga agggaatcct
     3061 gcaaaggtca acctgaaaag acctcctact gcagggtccc agttcaaggc gtcggtggcc
     3121 accctgatga aaaacctgca gaccaagaac ccgaactata tcaggtgcat caagccgaac
     3181 gataaaaagg ccgctcacat tttcagtgag agtctcgtgt gccatcagat ccggtacctg
     3241 ggtcttctgg agaacgttcg agtgagacgg gcaggctatg ccttcaggca ggcctatgaa
     3301 ccttgtctgg aaagatacaa gatgctttgc aaacaaacat ggcctcactg gaagggacca
     3361 gcaaggtctg gtgtggaagt tctgtttaat gagctagaga ttcccgtgga agagtactcc
     3421 ttcggtagat ccaagatatt catccgaaac ccaaggacat tattccaatt agaagaccta
     3481 aggaagcagc ggctggagga cctggccact ctcattcaga agatttatcg aggatggaaa
     3541 tgtcgtacac attttctgct catgaaaaga agccaagtcg tgattgccgc ctggtacagg
     3601 agatacgcgc aacaaaagcg atatcagcag ataaagagtt cagccctggt gattcagtct
     3661 tacatccggg gctggaaggc tagaaaaatc ctgcgggagc tgaagcatca gaaacgctgc
     3721 aaggaggcag ccaccaccat cgcagcatac tggcacggca cccaggcacg aagggaattg
     3781 aaacgcttga aggaggaggc taggcgtaag catgcagttg ctgtcatttg ggcttactgg
     3841 cttggactga aggtacgaag ggagtacagg aaattcttta gagcaaatgc tggaaagaaa
     3901 atctatgagt ttacacttca aagaattgtg caaaaatact tgttggaaat gaaaaataaa
     3961 atgccttcct tgtcaccaat agataaaaat tggccatcaa gaccttacct attcttggat
     4021 tcgactcaca aggagctaaa aaggatcttc cacttgtgga ggtgtaaaaa atacagggat
     4081 caattcacag accagcagaa acttatttat gaagagaagc tggaggccag tgaactcttc
     4141 aaagacaaga aggctttata tccttctagt gttggtcagc cattccaagg agcttacctg
     4201 gaaatcaaca agaacccaaa atataagaaa ctcaaagatg ccattgaaga gaagatcatt
     4261 attgctgagg ttgtgaacaa aatcaaccgg gctaatggga agagtacatc tcggattttc
     4321 ctcttaacaa acaataacct tctccttgct gaccaaaagt cggggcagat caagtccgag
     4381 gtccccctgg tggatgtgac taaggtctcc atgagttcac agaatgatgg tttctttgca
     4441 gtacacctca aggagggttc agaagcagct agtaagggtg actttctctt cagcagtgac
     4501 cacctgattg aaatggcaac caagttgtat cgcacgactc ttagccaaac taaacagaag
     4561 ctcaatatcg agatttccga tgagttcctg gtacagttca gacaggacaa agtatgtgtg
     4621 aagtttattc aaggcaacca gaaaaacggg agcgtgccaa cctgcaaacg aaagaacaac
     4681 aggcttcttg aagtcgctgt cccttaagtg aggatcaagc ttatcgatac cgtcgacggt
     4741 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     4801 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4861 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     5041 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5101 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5161 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5221 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5281 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5341 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5401 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5461 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5581 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5641 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5701 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5761 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5941 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6001 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6061 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6121 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6181 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6241 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6301 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6361 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6421 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6481 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6541 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6601 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6661 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6721 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6781 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6841 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6901 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6961 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7021 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7081 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7141 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7201 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7381 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7501 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7741 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7801 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7861 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7921 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7981 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8041 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
