LOCUS       sequence-267839-        7869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of rat Myo 1d (Myr 4) in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7869)
  AUTHORS   Bahler M, Kroschewski R, Stoffler HE, Behrmann T
  TITLE     Rat myr 4 defines a novel subclass of myosin I: identification, 
            distribution, localization, and mapping of calmodulin-binding sites 
            with differential calcium sensitivity.
  JOURNAL   J Cell Biol. 1994 Jul;126(2):375-89. doi: 10.1083/jcb.126.2.375.
  PUBMED    8034741
REFERENCE   2  (bases 1 to 7869)
  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"; doi: "10.1083/jcb.126.2";
            journalName: "J Cell Biol"; date: "1994-07"; volume: "126"; issue: 
            "2"; pages: "375-89"
COMMENT     Sequence Label: GFP-Myo 1d
FEATURES             Location/Qualifiers
     source          1..7869
                     /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    4657..4778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4694..4713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4748..4767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4785..5240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4922..4943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5134..5153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5267..5371
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5373..5730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5373..5564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5396..5416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5581..5716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5765..6559
                     /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             5765..6559
                     /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(5819..5838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6429..6448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6747..6766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6791..6838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7167..7755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7656..7675
                     /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 atctcgaatg gcggagcagg agagccttga gtttggcaag
     1381 gcggacttcg tgctgatgga caccgtctcc atgcccgagt tcatggccaa ccttaggctc
     1441 agatttgaga aaggacgcat ctatacattc attggcgagg tggtcgtctc cgtgaaccca
     1501 tacaaggtgc tgaacatcta tggaagggac acaattgagc agtacaaagg tcgggagctg
     1561 tacgagagac ctcctcatct cttcgctatt gctgacgctg cttacaaggc tatgaagagg
     1621 cgatcaaaag acacctgtat tatgatatca ggggaaagtg gagctggtaa aacagaggcc
     1681 agtaagtaca tcatgcagta tatcgccgcc atcaccaacc ccagccagag agcagagata
     1741 gaaagagtga agaatatgtt gctgaagtca aactgtgttt tggaagcttt tggaaatgcc
     1801 aaaaccaatc gcaatgacaa ttccagcagg tttgggaaat acatggatat caactttgac
     1861 ttcaagggag accccattgg gggacatatt aataactact tgctggaaaa gtctcgtgta
     1921 attgtgcaac agcccggaga acgaagcttc cattctttct atcagctact tcagggaggt
     1981 tccgagcaga tgctgcattc tctacatctg cagaaatcac tctcatccta caactacatc
     2041 cgtgtggggg cacaactgaa gtcttccatc aatgatgctg cagagttcaa agttgtagct
     2101 gatgccatga aagtaattgg attcaaacct gaggagattc aaacagtgta taaaattctg
     2161 gctgctattc ttcacttggg aaatttaaaa tttatagtag atggtgacac gccacttatt
     2221 gagaatggca aagttgtatc tgtcatagca gaattgctct caaccaaggc agacatggtg
     2281 gagaaagccc tgctttaccg gacggtggcc acaggccgtg acatcattga caaacagcac
     2341 acggaacagg aggctagcta cggcagagac gcctttgcca aggcaatata tgagcgcctc
     2401 ttttgttgga ttgttactcg catcaatgac atcatcgagg tcaagaacta cgacaccaca
     2461 gtacatggga aaaacacagt tattggcgtc ttggatatct atggctttga aatctttgac
     2521 aacaacagct tcgagcagtt ctgcattaac tactgcaatg agaagctgca gcagctgttc
     2581 attcagctgg tgctgaagca ggagcaagag gagtaccagc gggaaggaat cccctggaaa
     2641 cacattgatt acttcaacaa ccagatcatt gtggacctcg tggagcagca gcacaaaggc
     2701 atcattgcaa tcctggatga cgcgtgtatg aatgttggca aagtcactga tggaatgttc
     2761 ctcgaggccc ttaacagtaa attgggcaaa catggtcatt tttccagccg aaagacctgt
     2821 gcctcagaca aaatcctgga gtttgatcgg gactttcgaa tccggcatta tgcaggtgat
     2881 gtcgtgtatt ctgtcatcgg tttcattgac aaaaacaaag atactttatt tcaagacttc
     2941 aagcgcctca tgtataacag ctcaaatcct gtgctgaaga acatgtggcc ggaaggcaaa
     3001 ctgagcatta cagaggtgac caagaggcct ctgaccgccg ccaccctgtt taagaattcc
     3061 atgattgctt tagtagacaa ccttgcttca aaggagccat actatgtacg ttgcatcaaa
     3121 cccaacgaca agaagtcccc acagattttt gatgatgagc gctgccgcca tcaggtagaa
     3181 tacctgggac tcctggaaaa tgtgagagtg cgccgggcag ggtttgcctt ccgccagacc
     3241 tatgagaagt ttcttcacag atacaagatg atctctgagt tcacctggcc taaccacgac
     3301 cttccttcag acaaggaggc tgtcaagaaa ctcatagagc gatgtggctt tcaggatgac
     3361 gtagcatatg ggaagaccaa aattttcatt cgaacacccc gtacgctgtt cactttggaa
     3421 gaactccgcg cccagatgct cgtcagggtt gtcctcttcc tacaaaaggt gtggcgtggc
     3481 accctggccc gcatgcggta caagaggacc aaggcagctc tgacaataat caggtactac
     3541 cggcgctata aagtgaagtc gtacatccac gaggtagcca ggcgattcca cggggtcaag
     3601 aacatgagag actacgggaa gcacgtgaag tggccaaccc cacccaaagt cctgcggcgt
     3661 tttgaggagg ccctgcagtc gatttttaat agatggagag catcccaact catcaagacc
     3721 atacctgcat cagaccttcc ccaggtcagg gcaaaggttg cagccatgga gatgttgaaa
     3781 ggtcaaaggg cagaccttgg gctccagagg gcctgggaag gcaactatct tgcttcaaag
     3841 ccagatacac ctcagacctc gggcactttc gtcccagttg ctaatgagct gaaacgcaag
     3901 gacaaatata tgaatgttct cttttcctgt catgtccgaa aggtgaatcg atttagtaag
     3961 gtagaagacc gagcaatttt tgtcactgac cgtcacctgt ataaaatgga ccccactaag
     4021 cagtacaagg tgatgaagac catcccactg tacaatttga ctggcctgag tgtgtccaat
     4081 ggaaaggacc agcttgtagt gttccacaca aaagacaaca aagacctcat cgtctgcctc
     4141 ttcagcaaac agccgaccca tgagagccgg attggagaac ttgttggagt cctggtgaat
     4201 catttcaaga gtgagaagcg ccaccttcaa gtgaacgtca ccaacccggt gcagtgcagc
     4261 ctgcatggga agaaatgcac cgtatccgtg gagacccggc ttaaccagcc acagcctgac
     4321 ttcaccaaga accgctccgg cttcatcctc agtgtgcccg ggaattgact gctgcacacc
     4381 ggaggcctgg cccaggggag caagagcctg ggcctggcac acagcaggcc aggtcctcgg
     4441 cctcgccatc cccacatctg ttccacctcg ctcaacatcc caggagctca agcttcgaat
     4501 tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
     4561 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4621 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4681 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4741 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     4801 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4861 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4921 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4981 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5041 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5101 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5161 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5221 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5401 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5461 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5521 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5581 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5641 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5701 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5761 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5821 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5881 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5941 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6001 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     6061 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     6121 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6181 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6241 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     6301 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6361 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6421 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6481 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6541 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6601 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6661 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6721 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6781 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6841 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6901 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6961 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     7021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7801 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7861 gccatgcat
//
