LOCUS       sequence-410675-        7003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hARFGAP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7003)
  AUTHORS   Saitoh A, Shin HW, Yamada A, Waguri S, Nakayama K
  TITLE     Three homologous ArfGAPs participate in coat protein I-mediated 
            transport.
  JOURNAL   J Biol Chem. 2009 May 15;284(20):13948-13957. doi: 
            10.1074/jbc.M900749200. Epub 2009 Mar 19.
  PUBMED    19299515
REFERENCE   2  (bases 1 to 7003)
  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.M900749200"; journalName: "J Biol Chem"; date: 
            "2009-05-15- 15"; volume: "284"; issue: "20"; pages: "13948-13957"
COMMENT     Sequence Label: pcDNA3-hARFGAP3-HA
FEATURES             Location/Qualifiers
     source          1..7003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..28)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        200..579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        580..783
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     734..754
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     828..847
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        828..846
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2466..2492
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2495..2511)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2495..2511)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2521..2539)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2522..2539)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2559..2576)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2565..2789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2835..3263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2922..2941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3132..3153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3272..3292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3277..3606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3457..3592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3519..3538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3673..4467
                     /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(3727..3746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4337..4356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4641..4762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4678..4697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4732..4751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4811..4827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4811..4827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4824..4846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4835..4851
                     /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(4859..4865,4866..4883,4884..4889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4904..4925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5042..5059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5213..5801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5293..5312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5972..6763,6764..6832))
                     /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     6595..6614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6833..6937)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
       61 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
      121 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
      181 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      241 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      301 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      361 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      421 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      481 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      541 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      601 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      661 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      721 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      781 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
      841 tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc
      901 tggaattcac catgggggac cccagcaagc aggacatctt gaccatcttc aagcgcctcc
      961 gctcggtgcc cactaacaag gtgtgttttg attgtggtgc caaaaatccc agctgggcaa
     1021 gcataaccta tggagtgttc ctttgcattg attgctcagg gtcccaccgg tcacttggtg
     1081 ttcacttgag ttttattcga tctacagagt tggattccaa ctggtcatgg tttcagttgc
     1141 gatgcatgca agtcggagga aacgctagtg catcttcctt ttttcatcaa catgggtgtt
     1201 ccaccaatga caccaatgcc aagtacaaca gtcgtgctgc tcagctctat agggagaaaa
     1261 tcaaatcgct cgcctctcaa gcaacacgga agcatggcac tgatctgtgg cttgatagtt
     1321 gtgtggttcc acctttgtcc cctccaccaa aggaggaaga tttttttgcc tctcacgttt
     1381 ctcctgaggt gagtgacaca gcgtgggcat cagcaatagc agaaccatct tctttaacat
     1441 caaggcctgt ggaaaccact ttggaaaata atgaaggtgg acaagagcaa ggaccaagtg
     1501 tggaaggtct taatgtacca acaaaggcta ctttagaggt atcctctatc ataaaaaaga
     1561 aaccaaatca agctaaaaaa ggccttgggg ccaaaaaagg aagtttggga gctcagaaac
     1621 tggcaaacac atgctttaat gaaattgaaa aacaagctca agctgcggat aaaatgaagg
     1681 agcaggaaga cctggccaag gtggtatcta aagaagaatc aattgtttca tcattacgat
     1741 tagcctataa ggatcttgaa attcaaatga agaaagacga aaagatgaac attagtggca
     1801 aaaaaaatgt tgactcagac agactcggca tgggatttgg aaattgcaga agtgttattt
     1861 cacattcagt gacttcagat atgcagacca tagagcagga atcacccatt atggcaaaac
     1921 caagaaaaaa gtataatgat gacagtgacg attcatattt tacttccagc tcaaggtact
     1981 ttgacgagcc agtggagtta aggagcagtt ctttctctag ctgggatgac agttcagatt
     2041 cctattggaa aaaagagacc agcaaagata ctgaaacagt tctgaaaacc acaggctatt
     2101 cagacagacc tactgctcgc cgcaagccag attatgagcc agttgaaaat acagatgagg
     2161 cccagaagaa gtttggcaat gtcaaggcca tttcatcaga tatgtatttt ggaagacaat
     2221 cccaggctga ttatgagacc agggcccgcc tagagaggct gtcagcaagt tcctccataa
     2281 gctcggctga tctgttcgag gagccgagga agcagccagc agggaactac agcctgtcca
     2341 gcgtgctgcc caacgccccc gacatggcgc agttcaagca gggagtgaga tcggttgctg
     2401 gaaaactctc cgtctttgct aatggagtcg tgacttcaat tcaggatcgc tacggttctc
     2461 tcgagtatcc atacgatgta ccagactacg cagagatcca ctagttctag agggccctat
     2521 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     2581 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2641 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2701 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2761 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2821 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3181 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3241 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3541 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3601 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3661 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3721 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3781 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3841 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3901 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3961 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4021 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4081 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4141 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4201 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4261 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4321 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4381 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4441 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4621 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4741 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6961 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgc
//
