LOCUS       sequence-227205-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RalGDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  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 6422)
  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: 
            "10.1128/MCB.00242-10"; journalName: "Mol Cell Biol"; date: 
            "2010-08"; volume: "30"; issue: "16"; pages: "3956-69"
COMMENT     Sequence Label: GFP-RalGDS
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        141..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     769..788
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        769..787
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(822..824,825..827,828..1538)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(822..824,825..827,828..1538)
                     /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(867..888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1128..1147)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1475..1496
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1881..1899)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1882..1899)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1919..1936)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1925..2149
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2195..2623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2282..2301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2492..2513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2632..2652)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2637..2966
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2817..2952
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2879..2898
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3033..3827
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3087..3106)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3697..3716
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4001..4122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4038..4057)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4092..4111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4171..4187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4171..4187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4184..4206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4195..4211
                     /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(4219..4225,4226..4243,4244..4249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4264..4285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4402..4419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4573..5161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4653..4672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5332..6123,6124..6192))
                     /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     5955..5974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6193..6297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6372..6391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
       61 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      121 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      781 ctatagggag acccaagctt ggtaccgagc tcggatccac catggtgagc aagggcgagg
      841 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      901 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      961 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1021 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1081 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1141 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1201 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1261 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1321 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1381 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1441 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1501 ccgccgggat cactctcggc atggacgagc tgtacaaggg gaattcggca cgagccgcgc
     1561 tgccgctcta caaccagcag gtgggcgact gctgtatcat ccgcgtcagc ctggacgtgg
     1621 acaatggcaa catgtacaag agcatcctgg tgaccagcca agataaggct ccggctgtaa
     1681 tccgcaaggc catggacaaa cacaacctgg aggaggagga gccggaggac tatgagctgc
     1741 tgcagattct ctcagatgac cggaagctga agatccctga aaacgccaac gtcttctatg
     1801 ccatgaactc taccgccaac tatgactttg tcctcaagaa gcggacgggt cgactcgagc
     1861 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     1921 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     1981 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2041 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2101 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2161 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2221 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2281 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2341 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2401 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2461 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2521 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2581 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2641 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2701 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2761 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2821 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2881 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     2941 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3001 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3061 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3121 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3181 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3241 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3301 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3361 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3421 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3481 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3541 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3601 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3661 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3721 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3781 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3841 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3901 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3961 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4021 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4081 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4141 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4321 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4381 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4981 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5221 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5281 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5341 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5401 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5461 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5521 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5581 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5641 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5701 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5761 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5821 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5881 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5941 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6001 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6061 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6121 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6181 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6301 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6361 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     6421 tg
//
