LOCUS       sequence-227747-        5272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of bovine Rap1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5272)
  AUTHORS   Takahashi M, Dillon TJ, Liu C, Kariya Y, Wang Z, Stork PJ
  TITLE     Protein kinase A-dependent phosphorylation of Rap1 regulates its 
            membrane localization and cell migration.
  JOURNAL   J Biol Chem. 2013 Sep 27;288(39):27712-23. doi: 
            10.1074/jbc.M113.466904. Epub 2013 Aug 14.
  PUBMED    23946483
REFERENCE   2  (bases 1 to 5272)
  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"; doi: 
            "10.1074/jbc.M113.466904"; journalName: "J Biol Chem"; date: 
            "2013-09-27- 27"; volume: "288"; issue: "39"; pages: "27712-23"
COMMENT     Sequence Label: GFP-Rap1b S179AS180A
FEATURES             Location/Qualifiers
     source          1..5272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(634..636,637..639,640..1350)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(634..636,637..639,640..1350)
                     /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(679..700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(940..959)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1287..1308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2081..2202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2118..2137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2172..2191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2209..2664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2558..2577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2691..2795
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2797..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2797..2988
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2820..2840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3189..3983
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3189..3983
                     /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(3243..3262)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3853..3872
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4171..4190)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4215..4262
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4591..5179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5080..5099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
      661 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      721 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      781 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
      841 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      901 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      961 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1021 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1081 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1141 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1201 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1261 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1321 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     1381 aattctcgtg agtataagct agtcgttctt ggctctggag gcgttggaaa atctgctctg
     1441 actgtacagt ttgttcaagg aatttttgtt gaaaaatatg atcctacgat agaagattct
     1501 tatagaaagc aagttgaagt agatgcacag cagtgtatgc ttgaaatctt ggatactgca
     1561 ggaacggaac aattcacagc aatgagggat ttgtacatga aaaatggaca aggctttgcg
     1621 ttagtttatt ccatcacagc acagtccaca tttaatgatt tacaagatct gagagaacag
     1681 atacttcgag ttaaagacac ggatgatgtt ccaatgattc tggttggtaa taagtgtgac
     1741 ttggaagatg aaagagttgt aggaaaggaa caaggtcaaa atctagcaag acagtggaac
     1801 aactgtgcat tcttagaatc ctctgcaaaa tcaaaaataa atgttaatga gatcttttat
     1861 gacctagtgc gacagattaa cagaaaaact ccagtgcctg ggaaggcccg caaaaaggcg
     1921 gcatgtcagc tgctttaatc tagagggccc gggatccacc ggatctagat aactgatcat
     1981 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2041 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2161 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2221 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2281 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2341 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2401 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2461 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2521 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2581 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2641 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2701 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2761 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2821 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3181 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3241 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3301 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3361 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3421 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3481 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3541 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3601 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3661 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3721 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3781 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3841 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3901 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3961 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4021 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4081 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4141 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4201 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4261 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4321 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4381 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4441 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4501 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4561 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4621 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4681 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4741 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4801 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4861 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4921 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4981 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5041 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5101 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5161 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5221 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg at
//
