LOCUS       Exported                7976 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Full length Anillin rendered RNAi-resistant) in mammalian expression
            vector pEGFP-C1.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-Anillin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  AUTHORS   Piekny AJ, Glotzer M
  TITLE     Anillin is a scaffold protein that links RhoA, actin, and myosin 
            during cytokinesis.
  JOURNAL   Curr Biol. 2008 Jan 8;18(1):30-6. Epub 2007 Dec 27.
  PUBMED    18158243
REFERENCE   2  (bases 1 to 7976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7976
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      661..670
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             667..1383
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             667..1383
                     /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(712..733)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(973..992)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1320..1341
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4818..4939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4855..4874)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4909..4928
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4946..5401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5083..5104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5295..5314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5428..5532
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5534..5891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5557..5577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5742..5877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5804..5823
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5926..6720
                     /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(5980..5999)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6590..6609
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6908..6927)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6952..6999
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7328..7916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7817..7836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      661 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      721 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      781 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      841 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      901 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      961 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1021 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1081 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1141 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1201 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1261 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1321 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1381 aagtccggac tcagatctcg agctcaagct tcaatggatc cgtttacgga gaaactgctg
     1441 gagcgaaccc gtgccaggcg agagaatctt cagagaaaaa tggctgagag gcccacagca
     1501 gctccaaggt ctatgactca tgctaagcga gctagacagc cactttcaga agcaagtaac
     1561 cagcagcccc tctctggtgg tgaagagaaa tcttgtacaa aaccatcgcc atcaaaaaaa
     1621 cgctgttctg acaacactga agtagaagtt tctaacttgg aaaataaaca accagttgag
     1681 tcgacatctg caaaatcttg ttctccaagt cctgtgtctc ctcaggtgca gccacaagca
     1741 gcagatacca tcagtgattc tgttgctgtc ccggcatcac tgctgggcat gaggagaggg
     1801 ctgaactcaa gattggaagc aactgcagcc tcctcagtta aaacacgtat gcaaaaactt
     1861 gcagagcaac ggcgccgttg ggataatgat gatatgacag atgacattcc tgaaagctca
     1921 ctcttctcac caatgccatc agaggaaaag gctgcttccc ctcccagacc tctgctttca
     1981 aatgcctcgg caactccagt tggcagaagg ggccgtctgg ccaatcttgc tgcaactatt
     2041 tgctcctggg aagatgatgt aaatcactca tttgcaaaac aaaacagtgt acaagaacag
     2101 cctggtaccg cttgtttatc caaattttcc tctgcaagtg gagcatctgc taggatcaat
     2161 agcagcagtg ttaagcagga agctacattc tgttcccaaa gggatggcga cgcaagctta
     2221 aacaaggccc tatcctcaag tgctgctgat gcgtctttgg ttaatgcctc aatttccagc
     2281 tctgtgaaag ctacttctcc agtgaaatct actacatcta tcactgatgc taaaagttgt
     2341 gagggacaaa atcctgagct acttccaaaa actcctatta gtcctctgaa aacgggggta
     2401 tcgaaaccaa ttgtgaagtc aactttatcc cagacagttc catccaaggg agaattaagt
     2461 agagaaattt gtctgcaatc tcaatctaaa gacaaatcta cgacaccagg aggaacagga
     2521 attaagcctt tcctggaacg ctttggagag cgttgtcaag aacatagcaa agaaagtcca
     2581 gctcgtagca caccccacag aacccccatt attactccaa atacaaaggc catccaagaa
     2641 aaattattca agcaagacac atcttcatct actacccatt tagcacaaca gctcaagcag
     2701 gaacgtcaaa aagaactagc atgtcttcgt ggccgatttg acaagggcaa tatatggagt
     2761 gcagaaaaag gcggaaactc aaaaagcaaa caactagaaa ccaaacagga aactcactgt
     2821 cagagcactc ccctcaaaaa acaccaaggt gtttcaaaaa ctcagtcact tccagtaaca
     2881 gaaaaggtga ccgaaaacca gataccagcc aaaaattcta gtacagaacc taaagaagtg
     2941 atacgtgaaa ttgagatgag tgtggatgat gatgatatca atagttcgaa agtaattaat
     3001 gacctcttca gtgatgtcct agaggaaggt gaactagata tggagaagag ccaagaggag
     3061 atggatcaag cattagcaga aagcagcgaa gaacaggaag atgcactgaa tatctcctca
     3121 atgtctttac ttgcaccatt ggcacaaaca gttggtgtgg taagtccaga gagtttagtg
     3181 tccacaccta gactggaatt gaaagacacc agcagaagtg atgaaagtcc aaaaccagga
     3241 aaattccaaa gaactcgtgt ccctcgagct gaatctggtg atagccttgg ttctgaagat
     3301 cgtgatcttc tttacagcat tgatgcatat agatctcaaa gattcaaaga aacagaacgt
     3361 ccatcaataa agcaggtgat tgttcggaag gaagatgtta cttcaaaact ggatgaaaaa
     3421 aataatgcct ttccttgtca agttaatatc aaacagaaaa tgcaggaact caataacgaa
     3481 ataaatatgc aacagacagt gatctatcaa gctagccagg ctcttaactg ctgtgttgat
     3541 gaagaacatg gaaaagggtc cctagaagaa gctgaagcag aaagacttct tctaattgca
     3601 actgggaaga gaacactttt gattgatgaa ttgaataaat tgaagaacga aggacctcag
     3661 aggaagaata aggctagtcc ccaaagtgaa tttatgccat ccaaaggatc agttactttg
     3721 tcagaaatcc gcttgcctct aaaagcagat tttgtctgca gtacggttca gaaaccagat
     3781 gcagcaaatt actattactt aattatacta aaagcaggag ctgaaaatat ggtagccaca
     3841 ccattagcaa gtacttcaaa ctctcttaac ggtgatgctc tgacattcac tactacattt
     3901 actctgcaag atgtatccaa tgactttgaa ataaatattg aagtttacag cttggtgcaa
     3961 aagaaagatc cctcaggcct tgataagaag aaaaaaacat ccaagtccaa ggctattact
     4021 ccaaagcgac tcctcacatc tataaccaca aaaagcaaca ttcattcttc agtcatggcc
     4081 agtccaggag gtcttagtgc tgtgcgaacc agcaacttcg cccttgttgg atcttacaca
     4141 ttatcattgt cttcagtagg aaatactaag tttgttctgg acaaggtccc ctttttatct
     4201 tctttggaag gtcatattta tttaaaaata aaatgtcaag tgaattccag tgttgaagaa
     4261 agaggttttc taaccatatt tgaagatgtt agtggttttg gtgcctggca tcgaagatgg
     4321 tgtgttcttt ctggaaactg tatatcttat tggacttatc cagatgatga gaaacgcaag
     4381 aatcccatag gaaggataaa tctggctaat tgtaccagtc gtcagataga accagccaac
     4441 agagaatttt gtgcaagacg caacactttt gaattaatta ctgtccgacc acaaagagaa
     4501 gatgaccgag agactcttgt cagccaatgc agggacacac tctgtgttac caagaactgg
     4561 ctgtctgcag atactaaaga agagcgggat ctctggatgc aaaaactcaa tcaagttctt
     4621 gttgatattc gcctctggca acctgatgct tgctacaaac ctattggaaa gccttgaccg
     4681 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     4741 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4801 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4861 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4921 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5521 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5821 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5881 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5941 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6001 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6061 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6121 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6181 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6241 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6301 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6361 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6421 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6481 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6541 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6601 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6661 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6721 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6781 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6841 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6901 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6961 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7021 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7081 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7141 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7441 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7681 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatg
//
