LOCUS       Exported                8137 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mouse myosin-1e fused with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    Myo1E-pEGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8137)
  AUTHORS   Zhang Y, Cao F, Zhou Y, Feng Z, Sit B, Krendel M, Yu CH
  TITLE     Tail domains of myosin-1e regulate phosphatidylinositol signaling 
            and F-actin polymerization at the ventral layer of podosomes.
  JOURNAL   Mol Biol Cell. 2019 Mar 1;30(5):622-635. doi: 
            10.1091/mbc.E18-06-0398. Epub 2019 Jan 2.
  PUBMED    30601698
REFERENCE   2  (bases 1 to 8137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8137
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1316..2032
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1316..2032
                     /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(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5462..5478)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5462..5478)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    5628..5749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5665..5684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5719..5738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5756..6211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5893..5914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6105..6124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6238..6342
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6344..6701
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(6367..6387)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6552..6687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6614..6633
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6736..7530
                     /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(6790..6809)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7400..7419
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7718..7737)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7762..7809
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatctcga gctggacatg gtacaggatc aactggtagt ggaagtagta tgggcagcaa
     2101 aggcgcctac cgataccatt ggcagagcca caatgtcaag cacagtggtg tggacgacat
     2161 ggttctactg tccaagatca cagagagctc cattgtagag aacctgaaga agagatacat
     2221 ggacgactat atttttacat atatagggtc tgtattaatc tctgtcaacc cattcaagca
     2281 aatgccatat tttggagaaa aagaaatcga aatgtaccaa ggagcggcac agtatgagaa
     2341 cccgcctcat atctatgccc ttgccgatag tatgtacaga aacatgatca tcgacagaga
     2401 gaaccagtgt gtcatcatca gcggtgaaag tggtgctgga aagacagtgg ctgccaagta
     2461 cattatgagc tatgtctcca gagtgtctgg aggaggcccc aaggtccagc atgtgaagga
     2521 catcatcctt caatccaacc cgctcctgga ggccttcgga aacgccaaga cagtgcgcaa
     2581 caacaactcc agccgatttg gaaaatactt tgaaatccag ttcagtccag gtggggaacc
     2641 agatggtggc aagatctcca acttccttct ggaaaagtcc agagtggtga tgcggaaccc
     2701 gggggaacgg agcttccaca tattctacca gctcattgag ggtgcctctc cagagcagaa
     2761 gcaaagcctt ggcatcacca gcatggacta ctattactac ctgagcctct cggggtccta
     2821 caaggtggat gacattgatg acaagcggga tttccaggaa accctgcacg ccatgaatgt
     2881 gattggaatc ttctcggaag agcagactct cgtgttgcag attgtggctg gaattctcca
     2941 cttgggaaac atcagcttca aagaagtggg caactatgcc gccgtggaga gtgaagaatt
     3001 tttggctttt cctgcgtatc tgctgggtat aaaccaagac cggttaaaag agaagctaac
     3061 gagccgccag atggacagta agtggggagg aaagtcagag tccatccatg tgaccctcaa
     3121 cgtggagcag gcctgctaca ccagggacgc gctcgccaag gccctgcacg cacgagtctt
     3181 tgatttcttg gtagattcta tcaataaagc catggagaag gaccacgagg agtacaacat
     3241 tggagtcctg gacatctatg gctttgaaat atttcagaaa aatggctttg agcagttctg
     3301 catcaatttc gttaacgaaa aattacagca gatttttatt gagctgacgc taaaagcaga
     3361 gcaggaagaa tacgttcagg agggaataag atggacaccc attgaatatt tcaataataa
     3421 aatcgtgtgt gacctcatag agagcaaagt caaccctcct gggattatga gcatcctgga
     3481 tgacgtgtgc gccaccatgc atgctgtggg tgagggggct gaccagactc tgctacagaa
     3541 gctccagatg cagatcggga gccatgagca cttcaacagc tggaaccaag gcttcatcat
     3601 tcaccactac gctgggaagg tttcctatga catggacggc ttctgcgaaa ggaaccggga
     3661 tgtgcttttc atggacctga ttgaactcat gcagagcagt gagctgcctt ttataaagtc
     3721 tttatttcct gaaaacctgc aagctgacaa gaaagggcgc ccaacgaccg caggaagcaa
     3781 aataaagaaa caagccaacg acctggtgag caccctgatg aaatgcacac cccactacat
     3841 tcgctgcata aaaccaaacg aaaccaagaa gcctaaagac tgggaggaga gcagagtgaa
     3901 gcatcaagtg gagtacctgg gtctcaaaga aaacatccga gtgagaagag ctggttatgc
     3961 ttatcggcgt gtcttccaga agtttctaca gaggtatgcc attctgacca aagctacctg
     4021 gcctgtgtgg agaggagatg agaaacaagg tgtcctccat ctactgcagt ctgtcaacat
     4081 ggacagcgac cagttccagc ttgggagaag taaagtgttc atcaaagccc cagagtctct
     4141 gtttcttcta gaagagatga gagagagaaa gtacgatggg tacgcacgtg tgatacagaa
     4201 aacatggagg aagttcgtgg cccgcaagaa atatgttcaa atgagagaag aagcctcaga
     4261 cctgctgttg aacaagaagg agaggaggag aaacagcatc aaccgaaact tcatcgggga
     4321 ctacatcggg atggaggagc gcccggaact gcagcagttt gtaggcaaga gggagaagat
     4381 tgactttgct gacacagtca ccaagtacga caggaggttc aagggtgtca agcgagacct
     4441 gctgctaacc cccaagtgct tgtacttaat tggaagggag aaggtgaagc aaggcccaga
     4501 caaaggtgtt gtgaaggaag tcctgaagcg gaggatcgag gtggaaagga tcctgtccgt
     4561 gtccctcagt accatgcagg atgacatctt cattctccac gagcaagaat atgacagttt
     4621 gctggagtcc gtctttaaaa cggagttcct gagcctctta gcaaagcgat acgaggaaaa
     4681 aacccagaag caactacctc tgaaattcag caataccctt gaactgaagt tgaaaaagga
     4741 gaactggggt ccctggagtg cagggggctc ccggcaagtg cagttccacc aaggctttgg
     4801 tgacctggct atcctcaagc ccagcaacaa agtgctgcag gtcagcatcg gacccgggct
     4861 gcccaagaac tcccgtccta ccagaagaaa caccgtcacc agcagagggt atccaggtgg
     4921 aactaagaac aattacccga tgagagctgc ccctgctccc ccaggatgcc atcaaaacgg
     4981 agtcatacga aaccagtttg tgcctcctcc ccatgccttt ggaaaccaga ggtccaatca
     5041 gaaaagcctg tacacctcta tggcccgccc acccttgcca aggcaacagt ccaccggctc
     5101 tgaccgattg tctcagacgc cagagagcct ggatttcctc aaggtcccgg accagggtgt
     5161 tgctggagtc cgaagacaaa cgagcagccg gcctccccca gcagggggca gaccaaagcc
     5221 tcaacccaag cctaagcctc aggtgccaca gtgcaaggcc ttgtatgcct atgatgctca
     5281 ggacacggac gaactcagct tcaacgccaa cgatattatt gatattatta aagaagaccc
     5341 ttctggctgg tggacgggta gactacgagg caagcagggc ctgtttccta acaactacgt
     5401 caccaagatc tgagtcgacg cgcgcaaggg cgaattccac attggtcgct gcagcccggg
     5461 ggatccacta gttctagagc ggccgcaccg cgggcccggg atccaccgga tctagataac
     5521 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5581 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5641 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5701 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     5761 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5821 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5881 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5941 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6001 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6061 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6121 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6181 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     6241 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6301 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     6361 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6421 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6481 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6541 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6601 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6661 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     6721 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6781 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6841 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6901 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6961 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     7021 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     7081 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     7141 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     7201 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     7261 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     7321 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     7381 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     7441 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7501 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     7561 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     7621 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     7681 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     7741 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     7801 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     7861 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     7921 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     7981 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     8041 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8101 gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
