LOCUS       Exported                5263 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of bovine Rap1b.
ACCESSION   .
VERSION     .
KEYWORDS    mCherry-Rap1b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5263)
  AUTHORS   Liu C, Takahashi M, Li Y, Dillon TJ, Kaech S, Stork PJ
  TITLE     The interaction of Epac1 and Ran promotes Rap1 activation at the 
            nuclear envelope.
  JOURNAL   Mol Cell Biol. 2010 Aug;30(16):3956-69. doi: 10.1128/MCB.00242-10. 
            Epub 2010 Jun 14.
  PUBMED    20547757
REFERENCE   2  (bases 1 to 5263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5263
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        149..452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        453..656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     607..627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      695..704
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             701..1408
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(846..864)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1114..1133
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2139..2260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2176..2195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2230..2249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2267..2722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2616..2635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2749..2853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2855..3212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2878..2898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3125..3144
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3247..4041
                     /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(3301..3320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3911..3930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4229..4248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4273..4320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4649..5237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5138..5157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
       61 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
      121 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      181 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      241 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      301 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      361 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      421 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      481 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      541 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      601 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      661 tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      721 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
      781 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      841 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
      901 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
      961 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1021 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     1081 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     1141 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     1201 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     1261 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     1321 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     1381 caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa
     1441 ttctcgtgag tataagctag tcgttcttgg ctctggaggc gttggaaaat ctgctctgac
     1501 tgtacagttt gttcaaggaa tttttgttga aaaatatgat cctacgatag aagattctta
     1561 tagaaagcaa gttgaagtag atgcacagca gtgtatgctt gaaatcttgg atactgcagg
     1621 aacggaacaa ttcacagcaa tgagggattt gtacatgaaa aatggacaag gctttgcgtt
     1681 agtttattcc atcacagcac agtccacatt taatgattta caagatctga gagaacagat
     1741 acttcgagtt aaagacacgg atgatgttcc aatgattctg gttggtaata agtgtgactt
     1801 ggaagatgaa agagttgtag gaaaggaaca aggtcaaaat ctagcaagac agtggaacaa
     1861 ctgtgcattc ttagaatcct ctgcaaaatc aaaaataaat gttaatgaga tcttttatga
     1921 cctagtgcga caaattaaca gaaaaactcc agtgcctggg aaggcccgca aaaagtcatc
     1981 atgtcagctg ctttaatcta gagggcccgg gatccaccgg atctagataa ctgatcataa
     2041 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2101 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2221 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2701 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2821 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3241 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3301 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3361 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3421 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3481 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3541 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3601 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3661 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3721 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3781 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3841 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3901 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3961 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4021 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4081 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4141 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4201 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4261 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4321 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4381 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4441 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acg
//
