LOCUS       sequence-319418-        8110 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of rat WT Myosin Vb with deletion of the 1435 
            N-terminal residues.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8110)
  AUTHORS   Frank M, Citarella CG, Quinones GB, Bentley M
  TITLE     A Novel Labeling Strategy Reveals That Myosin Va and Myosin Vb Bind 
            the Same Dendritically Polarized Vesicle Population.
  JOURNAL   Traffic. 2020 Sep 21. doi: 10.1111/tra.12764.
  PUBMED    32959500
REFERENCE   2  (bases 1 to 8110)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Traffic. 
            2020 Sep 21. doi: 10.1111/tra.12764."
COMMENT     Sequence Label: pBa.GFP-myosin Vb tail 1436-1846
FEATURES             Location/Qualifiers
     source          1..8110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        148..423
                     /label=chicken beta-actin promoter
     primer_bind     1366..1388
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             join(1456..1458,1459..1461,1462..2172)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGGELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1501..1522)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1762..1781)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2109..2130
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3676..3810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3713..3732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3767..3786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3841..3857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3841..3857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3854..3876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3865..3881
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3889..3895,3896..3913,3914..3919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3934..3955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4072..4089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4243..4831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4323..4342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5002..5793,5794..5862))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5625..5644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5863..5967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6035..6053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6091..6113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6213..6232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6426..6448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6440..6457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6441..6457
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcaagggga ccgcaggacc acctcggggg tggggggagg gctgcacacg cggaccccgc
       61 tccccctccc caacaaagca ctgtggaatc aaaaaggggg gaggggggat ggaggggcgc
      121 gtcacacccc cgccccacac cctcacctcg aggtgagccc cacgttctgc ttcactctcc
      181 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
      241 cagcgatggg ggsggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      301 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      361 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      421 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      481 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      541 cctccgggct gtaattagcg cttggtttaa tgacggctcg tttcttttct gtggctgcgt
      601 gaaagcctta aagggctccg ggagggccct ttgtgcgggg gggagcggct cggggggtgc
      661 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      721 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
      781 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
      841 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt aacccccccc
      901 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg
      961 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1021 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1081 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1141 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1201 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1261 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct
     1321 gtccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1381 gtgtgaccgg cggggtttat atcttccctt ctctgttcct ccgcagcccc caagcttggc
     1441 gcgcctaggc ctaccatggt gagcaagggc ggggagctgt tcaccggggt ggtgcccatc
     1501 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1561 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1621 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1681 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1741 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1801 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1861 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1921 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1981 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2041 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2161 gagctgtaca agtccggact cagatctcga gaagcccagg cattggcaca gagtgacagg
     2221 aggcaccatg aactcacaag acaggtcacg gtccaacgaa aagagaagga cttccaaggc
     2281 atgctggagt accacaaaga ggacgaggca ctccttatcc ggaacctggt gacagacctg
     2341 aagccccaga tgctgtcggg caccgtgccc tgtctgcctg catacatact ctacatgtgc
     2401 atcaggcacg cggactacac caacgatgac ctcaaggtgc actcgttgct gagctccacc
     2461 atcaatggca ttaagaaagt cctcaagaag cacaatgagg actttgagat gacgtcattc
     2521 tggttatcca acacctgccg cctccttcac tgtttgaagc agtacagtgg ggatgagggt
     2581 ttcatgacac agaacacggc gaagcagaat gagcactgtc tcaagaactt tgacctcact
     2641 gaataccgtc aggtgctaag tgacctttcc attcagatct atcagcaact cattaaaatt
     2701 gctgagggcc tgctacagcc tatgatagtt tctgccatgc tggaaaatga gagtatccag
     2761 gggctgtcag gggtgaggcc aactggctac cggaagcgct cgtccagcat ggtggacgga
     2821 gaaaattcgt attgcctgga ggccatcatc cgccagatga atttctttca tacagtcctg
     2881 tgtgaccagg gcctggaccc tgagattatc ctgcaggtgt tcaaacagct tttctacatg
     2941 atcaatgcgg tgactcttaa caacctactc ctgcggaaag atgcctgctc ctggagcacc
     3001 ggcatgcaac tcaggtacaa cataagtcaa ctcgaagagt ggcttcgggg aaaaaacctt
     3061 cagcagagtg gagcggttca gaccatggag cccttgatcc aggcagccca actccttcag
     3121 ctgaagaaga agacccagga ggatgctgag gccatctgct ctctgtgtac ctccctcagc
     3181 acccagcaga ttgtcaaaat tttaaacctc tacactccct tgaatgaatt tgaagaacgg
     3241 gtgacagtgt cctttatacg aacaatccag gctcagctac aagagaggag tgacccccag
     3301 cagctactgc tggactccaa acacatgttt cctgttctgt ttccatttaa tccatctgct
     3361 ctgaccatgg actcaatcca catcccggcc tgtctcaatc tggagttcct caatgaagtc
     3421 tgaggatgcg ttgtttcaag ccgaattctg cagatatcca gcacagtggc ggccgctcga
     3481 gtctagaggg cccgtttaaa cccgctgatc agcctcgagg ctgatcagcg ggtttaaacg
     3541 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     3601 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3661 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3721 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3781 ctcatcaatg tatcttatca tgtctggatc cccgggtacc gagctcgaat tcgtaatcat
     3841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6061 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6241 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6421 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagttgggat ctttgcattg
     6481 gcccacggct ctcaggatgg ggatgctccc cttcagcacc cggttcccct tggaaactga
     6541 tggtcctggc tctgtggcat ggcagtggca ctgtgaggag ccccctacca gcagcacaca
     6601 gtggggttgg cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg
     6661 aacccgaatt gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca
     6721 cgctgccttc tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca
     6781 tttcccattg ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg
     6841 ctgcccttct ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca
     6901 aacgttaatt aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga
     6961 gtggggctga tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc
     7021 ccacggaggt ccccatgtgc tgcagtgtca ccgcctccga ggagacagta ttgtccctgc
     7081 ggtgtccctg cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg
     7141 cagccccgat gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg
     7201 cacacagggc tgggtcccgc tcccctaata tctacagtgc ttcttgcatg gccccttaat
     7261 caatgcagtt aatcagcatg cgctcatgca ccgctctgga gctgcaaagc ccctcgcagc
     7321 gctgctcacc aacaccgcgc accgccccgg cccagcctgc agcacgcgct gcaaacagga
     7381 aagaaacaaa atattgccca aatgtaggca aaggcattcg gctgccttga cctccgccgg
     7441 gccgggccct gcctgactca gctccttact cagcgctcgc ttcctccctc cggctgccac
     7501 cgccgcagcg cacaccctga caaagagtgg cccttaacgg gctctgaggt gcacccagca
     7561 gtgcactcag cagtccaagg gccggcctgg aggtttgcac cgctacgtgc tgacattagc
     7621 attgaacttg gccctgggta gtgctgcagg ccgggcgggg tgggtgtaga gagtgcagcg
     7681 cgcgttgcac ccggtgcccc ttcccctccc ttgcatccca gcaggctgca ccccagcacc
     7741 aggcccgtgc atgcatgctc ctggtgttat tgcagcctgg tgcatgcatg cgtcttgatg
     7801 gtgcagcgct gtgcatgcat cctccttggt gtgtagcagt ctagtgcatg catacccctc
     7861 ggtgttattg ctgctctgtg cacgcacgct cattgtatca cttcatccca gtgcatgcac
     7921 tcacactgga gcgattgctg ctcggtgcac gcacactcat tgtatcactg cagctcagtg
     7981 gctgcacgca caccggtgtt attgctgctc ggtgcgtgca tgcacatcag tgtcgctgca
     8041 gctcagtgca tgcacgctca ttgcccatcg ctatccctgc ctctcctgct ggcgctcccc
     8101 gggaggtgac
//
