LOCUS       sequence-245057-        8096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FLAG-MYO1D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8096)
  AUTHORS   McAlpine W, Wang KW, Choi JH, San Miguel M, McAlpine SG, Russell J, 
            Ludwig S, Li X, Tang M, Zhan X, Choi M, Wang T, Bu CH, Murray AR, 
            Moresco EMY, Turer EE, Beutler B
  TITLE     The class I myosin MYO1D binds to lipid and protects against 
            colitis.
  JOURNAL   Dis Model Mech. 2018 Sep 27;11(9). pii: 11/9/dmm035923. doi: 
            10.1242/dmm.035923.
  PUBMED    30279225
REFERENCE   2  (bases 1 to 8096)
  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: "Dis Model 
            Mech."; date: "2018-09-27"; volume: "11(9). pii: 11/9/dmm035923. 
            doi"; pages: " 10.1242/dmm.035923"
COMMENT     Sequence Label: pcDNA-FLAG-MYO1D
FEATURES             Location/Qualifiers
     source          1..8096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             696..719
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3818..3835)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3824..4048
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4094..4522
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4181..4200)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4391..4412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4531..4551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4536..4865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4716..4851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4778..4797
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4913..4960
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4979..5377
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5535..5656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5572..5591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5626..5645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5705..5721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5705..5721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5718..5740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5729..5745
                     /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(5753..5759,5760..5777,5778..5783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5798..5819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5936..5953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6107..6695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6187..6206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6866..7657,7658..7726))
                     /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     7489..7508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7727..7831)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7906..7925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaga gctccttaag gttaacgcca ccatggacta caaagacgat gacgacaagt
      721 ctagagaatt cggatccgcg gagcaggaga gcctggaatt cggcaaggca gacttcgtgc
      781 tgatggacac cgtctccatg cccgagttca tggccaacct caggctcaga tttgaaaaag
      841 ggcgcatcta tacgttcatt ggagaagtcg tcgtttctgt gaacccttac aagttgttga
      901 acatctatgg aagagacaca attgagcagt ataaaggccg tgagctgtat gagagaccgc
      961 ctcacctttt tgctattgcg gatgctgctt acaaggctat gaagaggcga tcaaaagaca
     1021 cttgtattgt gatatcaggg gaaagtggag ctggtaaaac ggaagccagt aagtacatta
     1081 tgcagtatat tgcggccatc accaacccca gtcagagagc agaggttgaa agagtgaaga
     1141 atatgttgct taagtccaac tgtgttttgg aagcttttgg aaatgccaaa accaaccgta
     1201 atgacaactc aagcaggttt ggaaaataca tggatatcaa ctttgacttc aagggtgacc
     1261 ctattggtgg gcatatcaat aactacttac tagaaaagtc tcgagtgatt gtgcaacagc
     1321 caggagaaag aagctttcat tctttctatc agctactcca aggaggttca gaacaaatgc
     1381 tacgctctct acatctccag aaatcccttt catcctacaa ctatattcat gtgggagctc
     1441 aattaaagtc ttctatcaat gatgctgccg aattcagagt tgttgctgat gccatgaaag
     1501 tcattggctt caaacctgag gagatccaaa cagtgtataa gattttggct gctattctgc
     1561 acttgggaaa tttaaaattt gtagtagatg gtgacacgcc tcttattgag aatggcaaag
     1621 tagtatctat catagcagaa ttgctctcta ctaagacaga tatggttgag aaagcccttc
     1681 tttaccggac tgtggccaca ggccgtgaca tcattgacaa gcagcacaca gaacaagagg
     1741 ccagctacgg cagagacgcc tttgccaagg caatatatga gcgccttttt tgttggatcg
     1801 ttactcgcat caatgatatt attgaggtca agaactatga caccacaatc catgggaaaa
     1861 acactgttat tggtgtcttg gatatctatg gctttgaaat cttcgacaac aacagttttg
     1921 aacaattctg tatcaattac tgcaatgaga aactgcagca gctatttatt cagctggttc
     1981 tgaagcaaga acaagaggaa taccagcggg aagggatccc ctggaaacat attgactact
     2041 tcaacaatca gatcattgtt gacctcgtgg agcaacagca caaagggatc attgcaatcc
     2101 ttgatgatgc ttgcatgaat gtcggcaaag tcaccgatga aatgtttctt gaagcactta
     2161 acagtaaatt gggcaaacac gcccattttt ccagccgaaa gctctgtgcc tcagacaaaa
     2221 ttctggagtt tgatcgagat tttcgaattc gacattatgc aggcgatgta gtctattctg
     2281 tcattggttt tattgacaaa aataaagata ctttatttca agatttcaag cgccttatgt
     2341 ataacagttc aaatcctgtg ctcaagaata tgtggcctga aggcaaactg agcattacag
     2401 aggtgaccaa gcgacctctg actgctgcta ccttgtttaa gaattctatg attgctctag
     2461 tagacaacct tgcatcaaag gaaccatatt acgttcgttg catcaaaccc aatgacaaga
     2521 aatctccaca gatatttgat gatgaacgct gccggcacca agtagaatat cttggactac
     2581 tggaaaatgt gagagtgcgt cgggcaggat ttgccttccg ccagacatac gagaagtttc
     2641 ttcacaggta taagatgatc tctgaattca cctggcccaa ccatgacctt ccttcagaca
     2701 aagaggctgt caagaaacta attgaacggt gtggttttca ggatgatgta gcttatggga
     2761 agaccaaaat tttcattcga acaccccgaa cattgtttac cttggaagaa ctccgtgccc
     2821 agatgctcat aaggattgtc ctctttctac aaaaggtgtg gcggggcacc ctggcccgca
     2881 tgcggtacaa aagaaccaag gcagctctga caataatcag gtactaccgg cgctacaaag
     2941 tgaagtcgta catccacgag gtggccagac gcttccatgg cgtcaagacc atgcgagact
     3001 acgggaagca cgtgaagtgg ccaagccctc ctaaagttct tcgccgtttt gaggaggccc
     3061 tgcagacgat tttcaataga tggagagcat cccagctcat caagagcatt ccggcctcag
     3121 acctgcccca ggtcagggca aaggttgcag ccgtggaaat gttgaagggt caaagggctg
     3181 acctcgggct ccagagggcc tgggagggca actatcttgc ttcaaagcca gatacacctc
     3241 agacctcagg cacttttgtc cctgttgcta atgaattgaa acggaaggac aaatacatga
     3301 atgtcctctt ttcctgtcac gtccgtaagg taaatcgatt tagtaaggtg gaagacagag
     3361 caatttttgt cactgaccgt cacctgtata aaatggatcc cactaaacag tacaaggtga
     3421 tgaagactat ccctctatac aatttgactg gtctgagtgt ctccaatgga aaggaccaac
     3481 ttgtagtgtt ccatacgaaa gacaacaaag acctcattgt ctgcctcttc agcaaacagc
     3541 caacccatga gagtcgaatt ggagaacttg ttggagtgct ggtgaatcat ttcaagagtg
     3601 agaagcgcca ccttcaagtg aacgtcacca acccagtaca gtgcagcctg cacgggaaga
     3661 agtgcaccgt ctccgtggag acgcggctca accagcccca gcccgacttc accaagaatc
     3721 gctcgggctt catcctcagc gtgcccggga actgactcga gccatggaag cttgatatct
     3781 aactgactga accggtggta ccaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3841 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3901 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3961 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4021 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4081 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4141 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4201 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4261 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4321 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4381 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4441 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4501 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4621 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4681 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4741 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4801 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4861 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     4921 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     4981 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     5041 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     5101 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     5161 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     5221 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     5281 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     5341 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
     5401 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5461 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5521 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5581 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5641 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5701 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5761 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5821 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5941 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6001 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6061 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6121 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6181 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6241 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6301 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6361 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6421 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6481 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6541 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6601 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6661 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6721 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6781 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6841 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6901 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6961 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7021 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7081 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7141 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7201 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7261 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7321 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7381 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7441 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7501 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7561 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7621 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7681 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7741 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7801 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7861 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7921 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7981 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     8041 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
