LOCUS       sequence-239540-        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"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4052..4069)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4058..4282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4328..4756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4625..4646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4765..4785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4770..5099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4950..5085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5012..5031
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5147..5194
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5213..5611
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5769..5890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5806..5825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5860..5879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5939..5955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5939..5955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5952..5974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5963..5979
                     /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(5987..5993,5994..6011,6012..6017))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6032..6053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6170..6187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6341..6929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6421..6440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7100..7891,7892..7960))
                     /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     7723..7742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7961..8065)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 tagagctcct taaggttaac gccaccatgg actacaaaga cgatgacgac aagtctagag
      961 aattcggatc cgcggagcag gagagcctgg aattcggcaa ggcagacttc gtgctgatgg
     1021 acaccgtctc catgcccgag ttcatggcca acctcaggct cagatttgaa aaagggcgca
     1081 tctatacgtt cattggagaa gtcgtcgttt ctgtgaaccc ttacaagttg ttgaacatct
     1141 atggaagaga cacaattgag cagtataaag gccgtgagct gtatgagaga ccgcctcacc
     1201 tttttgctat tgcggatgct gcttacaagg ctatgaagag gcgatcaaaa gacacttgta
     1261 ttgtgatatc aggggaaagt ggagctggta aaacggaagc cagtaagtac attatgcagt
     1321 atattgcggc catcaccaac cccagtcaga gagcagaggt tgaaagagtg aagaatatgt
     1381 tgcttaagtc caactgtgtt ttggaagctt ttggaaatgc caaaaccaac cgtaatgaca
     1441 actcaagcag gtttggaaaa tacatggata tcaactttga cttcaagggt gaccctattg
     1501 gtgggcatat caataactac ttactagaaa agtctcgagt gattgtgcaa cagccaggag
     1561 aaagaagctt tcattctttc tatcagctac tccaaggagg ttcagaacaa atgctacgct
     1621 ctctacatct ccagaaatcc ctttcatcct acaactatat tcatgtggga gctcaattaa
     1681 agtcttctat caatgatgct gccgaattca gagttgttgc tgatgccatg aaagtcattg
     1741 gcttcaaacc tgaggagatc caaacagtgt ataagatttt ggctgctatt ctgcacttgg
     1801 gaaatttaaa atttgtagta gatggtgaca cgcctcttat tgagaatggc aaagtagtat
     1861 ctatcatagc agaattgctc tctactaaga cagatatggt tgagaaagcc cttctttacc
     1921 ggactgtggc cacaggccgt gacatcattg acaagcagca cacagaacaa gaggccagct
     1981 acggcagaga cgcctttgcc aaggcaatat atgagcgcct tttttgttgg atcgttactc
     2041 gcatcaatga tattattgag gtcaagaact atgacaccac aatccatggg aaaaacactg
     2101 ttattggtgt cttggatatc tatggctttg aaatcttcga caacaacagt tttgaacaat
     2161 tctgtatcaa ttactgcaat gagaaactgc agcagctatt tattcagctg gttctgaagc
     2221 aagaacaaga ggaataccag cgggaaggga tcccctggaa acatattgac tacttcaaca
     2281 atcagatcat tgttgacctc gtggagcaac agcacaaagg gatcattgca atccttgatg
     2341 atgcttgcat gaatgtcggc aaagtcaccg atgaaatgtt tcttgaagca cttaacagta
     2401 aattgggcaa acacgcccat ttttccagcc gaaagctctg tgcctcagac aaaattctgg
     2461 agtttgatcg agattttcga attcgacatt atgcaggcga tgtagtctat tctgtcattg
     2521 gttttattga caaaaataaa gatactttat ttcaagattt caagcgcctt atgtataaca
     2581 gttcaaatcc tgtgctcaag aatatgtggc ctgaaggcaa actgagcatt acagaggtga
     2641 ccaagcgacc tctgactgct gctaccttgt ttaagaattc tatgattgct ctagtagaca
     2701 accttgcatc aaaggaacca tattacgttc gttgcatcaa acccaatgac aagaaatctc
     2761 cacagatatt tgatgatgaa cgctgccggc accaagtaga atatcttgga ctactggaaa
     2821 atgtgagagt gcgtcgggca ggatttgcct tccgccagac atacgagaag tttcttcaca
     2881 ggtataagat gatctctgaa ttcacctggc ccaaccatga ccttccttca gacaaagagg
     2941 ctgtcaagaa actaattgaa cggtgtggtt ttcaggatga tgtagcttat gggaagacca
     3001 aaattttcat tcgaacaccc cgaacattgt ttaccttgga agaactccgt gcccagatgc
     3061 tcataaggat tgtcctcttt ctacaaaagg tgtggcgggg caccctggcc cgcatgcggt
     3121 acaaaagaac caaggcagct ctgacaataa tcaggtacta ccggcgctac aaagtgaagt
     3181 cgtacatcca cgaggtggcc agacgcttcc atggcgtcaa gaccatgcga gactacggga
     3241 agcacgtgaa gtggccaagc cctcctaaag ttcttcgccg ttttgaggag gccctgcaga
     3301 cgattttcaa tagatggaga gcatcccagc tcatcaagag cattccggcc tcagacctgc
     3361 cccaggtcag ggcaaaggtt gcagccgtgg aaatgttgaa gggtcaaagg gctgacctcg
     3421 ggctccagag ggcctgggag ggcaactatc ttgcttcaaa gccagataca cctcagacct
     3481 caggcacttt tgtccctgtt gctaatgaat tgaaacggaa ggacaaatac atgaatgtcc
     3541 tcttttcctg tcacgtccgt aaggtaaatc gatttagtaa ggtggaagac agagcaattt
     3601 ttgtcactga ccgtcacctg tataaaatgg atcccactaa acagtacaag gtgatgaaga
     3661 ctatccctct atacaatttg actggtctga gtgtctccaa tggaaaggac caacttgtag
     3721 tgttccatac gaaagacaac aaagacctca ttgtctgcct cttcagcaaa cagccaaccc
     3781 atgagagtcg aattggagaa cttgttggag tgctggtgaa tcatttcaag agtgagaagc
     3841 gccaccttca agtgaacgtc accaacccag tacagtgcag cctgcacggg aagaagtgca
     3901 ccgtctccgt ggagacgcgg ctcaaccagc cccagcccga cttcaccaag aatcgctcgg
     3961 gcttcatcct cagcgtgccc gggaactgac tcgagccatg gaagcttgat atctaactga
     4021 ctgaaccggt ggtaccaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4081 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4141 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4201 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4261 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4321 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4381 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4441 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4501 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4561 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4621 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4681 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4741 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4921 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4981 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5041 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5101 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     5161 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     5221 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     5281 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     5341 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     5401 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     5461 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     5521 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     5581 gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag caggactgac
     5641 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5701 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5761 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5881 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5941 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6001 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6061 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6781 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7381 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7681 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
