LOCUS       sequence-227668-        7122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of  rat myosin 1D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7122)
  AUTHORS   Saydmohammed M, Yagi H, Calderon M, Clark MJ, Feinstein T, Sun M, 
            Stolz DB, Watkins SC, Amack JD, Lo CW, Tsang M
  TITLE     Vertebrate myosin 1d regulates left-right organizer morphogenesis 
            and laterality.
  JOURNAL   Nat Commun. 2018 Aug 23;9(1):3381. doi: 10.1038/s41467-018-05866-2.
  PUBMED    30139971
REFERENCE   2  (bases 1 to 7122)
  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: "Nat 
            Commun."; date: "2018-08-23"; volume: "9(1):3381. doi"; pages: " 
            10.1038/s41467-018-05866-2"
COMMENT     Sequence Label: pCS2+ rat MYO1D
FEATURES             Location/Qualifiers
     source          1..7122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    3174..3308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3252..3271
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3421..3441)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3421..3439)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3460..3476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3460..3476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3473..3495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3484..3500
                     /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(3508..3514,3515..3532,3533..3538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3553..3574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3691..3708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3862..4450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3942..3961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4621..5412,5413..5481))
                     /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     5244..5263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5482..5586)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5612..6067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5749..5770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5961..5980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6138..7122
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg gcggagcagg agagccttga gtttggcaag
      121 gcggacttcg tgctgatgga caccgtctcc atgcccgagt tcatggccaa ccttaggctc
      181 agatttgaga aaggacgcat ctatacattc attggcgagg tggtcgtttc cgtgaaccca
      241 tacaaggtgc tgaacatcta tggaagggac acaattgagc agtacaaagg tcgggagctg
      301 tacgagagac ctcctcatct cttcgctatt gctgacgctg cttacaaggc tatgaagagg
      361 cgatcaaaag acacctgtat tatgatatca ggggaaagtg gagctggtaa aacagaggcc
      421 agtaagtaca tcatgcagta tatcgccgcc atcaccaacc ccagccagag agcagagata
      481 gaaagagtga agaatatgtt gctgaagtca aactgtgttt tggaagcttt tggaaatgcc
      541 aaaaccaatc gcaatgacaa ttccagcagg tttgggaaat acatggatat caactttgac
      601 ttcaagggag accccattgg gggacatatt aataactact tgctggaaaa gtctcgtgta
      661 attgtgcaac agcccggaga acgaagcttc cattctttct atcagctact tcagggaggt
      721 tccgagcaga tgctgcattc tctacatctg cagaaatcac tctcatccta caactacatc
      781 cgtgtggggg cacaactgaa gtcttccatc aatgatgctg cagagttcaa agttgtagct
      841 gatgccatga aagtaattgg attcaaacct gaggagattc aaacagtgta taaaattctg
      901 gctgctattc ttcacttggg aaatttaaaa tttatagtag atggtgacac gccacttatt
      961 gagaatggca aagttgtatc tgtcatagca gaattgctct caaccaaggc agacatggtg
     1021 gagaaagccc tgctttaccg gacggtggcc acaggccgtg acatcattga caaacagcac
     1081 acggaacagg aggctagcta cggcagagac gcctttgcca aggcaatata tgagcgcctc
     1141 ttttgttgga ttgttactcg catcaatgac atcatcgagg tcaagaacta cgacaccaca
     1201 gtacatggga aaaacacagt tattggcgtc ttggatatct atggctttga aatctttgac
     1261 aacaacagct tcgagcagtt ctgcattaac tactgcaatg agaagctgca gcagctgttc
     1321 attcagctgg tgctgaagca ggagcaagag gagtaccagc gggaaggaat cccctggaaa
     1381 cacattgatt acttcaacaa ccagatcatt gtggacctcg tggagcagca gcacaaaggc
     1441 atcattgcaa tcctggatga cgcgtgtatg aatgttggca aagtcactga tggaatgttc
     1501 ctcgaggccc ttaacagtaa attgggcaaa catggtcatt tttccagccg aaagacctgt
     1561 gcctcagaca aaatcctgga gtttgatcgg gactttcgaa tccggcatta tgcaggtgat
     1621 gtcgtgtatt ctgtcatcgg tttcattgac aaaaacaaag atactttatt tcaagacttc
     1681 aagcgcctca tgtataacag ctcaaatcct gtgctgaaga acatgtggcc ggaaggcaaa
     1741 ctgagcatta cagaggtgac caagaggcct ctgaccgccg ccaccctgtt taagaattcc
     1801 atgattgctt tagtagacaa ccttgcttca aaggagccat actatgtacg ttgcatcaaa
     1861 cccaacgaca agaagtcccc acagattttt gatgatgagc gctgccgcca tcaggtagaa
     1921 tacctgggac tcctggaaaa tgtgagagtg cgccgggcag ggtttgcctt ccgccagacc
     1981 tatgagaagt ttcttcacag atacaagatg atctctgagt tcacctggcc taaccacgac
     2041 cttccttcag acaaggaggc tgtcaagaaa ctcatagagc gatgtggctt tcaggatgac
     2101 gtagcatatg ggaagaccaa aattttcatt cgaacacccc gtacgctgtt cactttggaa
     2161 gaactccgcg cccagatgct cgtcagggtt gtcctcttcc tacaaaaggt gtggcgtggc
     2221 accctggccc gcatgcggta caagaggacc aaggcagctc tgacaataat caggtactac
     2281 cggcgctata aagtgaagtc gtacatccac gaggtagcca ggcgattcca cggggtcaag
     2341 aacatgagag actacgggaa gcacgtgaag tggccaaccc cacccaaagt cctgcggcgt
     2401 tttgaggagg ccctgcagtc gatttttaat agatggagag catcccaact catcaagacc
     2461 atacctgcat cagaccttcc ccaggtcagg gcaaaggttg cagccatgga gatgttgaaa
     2521 ggtcaaaggg cagaccttgg gctccagagg gcctgggaag gcaactatct tgcttcaaag
     2581 ccagatacac ctcagacctc gggcactttc gtcccagttg ctaatgagct gaaacgcaag
     2641 gacaaatata tgaatgttct cttttcctgt catgtccgaa aggtgaatcg atttagtaag
     2701 gtagaagacc gagcaatttt tgtcactgac cgtcacctgt ataaaatgga ccccactaag
     2761 cagtacaagg tgatgaagac catcccactg tacaatttga ctggcctgag tgtgtccaat
     2821 ggaaaggacc agcttgtagt gttccacaca aaagacaaca aagacctcat cgtctgcctc
     2881 ttcagcaaac agccgaccca tgagagccgg attggagaac ttgttggagt cctggtgaat
     2941 catttcaaga gtgagaagcg ccaccttcaa gtgaacgtca ccaacccggt gcagtgcagc
     3001 ctgcatggga agaaatgcac cgtatccgtg gagacccggc ttaaccagcc acagcctgac
     3061 ttcaccaaga accgctccgg cttcatcctc agtgtgcccg ggaattgagg atcccatcga
     3121 ttcgaattca aggcctctcg agcctctaga actatagtga gtcgtattac gtagatccag
     3181 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3241 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3301 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3361 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     3421 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3481 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     3541 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3601 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3661 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3721 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3781 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3841 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4261 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4441 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4501 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4561 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4681 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4801 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5041 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5281 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5461 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5521 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5581 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6061 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6121 tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat
     6181 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
     6241 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
     6301 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
     6361 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
     6421 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
     6481 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
     6541 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
     6601 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
     6661 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
     6721 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
     6781 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
     6841 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
     6901 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
     6961 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
     7021 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     7081 gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
