LOCUS       sequence-410130-        6673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hARFGAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6673)
  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 6673)
  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-hARFGAP1-HA
FEATURES             Location/Qualifiers
     source          1..6673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2171..2197
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2200..2216)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2200..2216)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2226..2244)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2227..2244)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2264..2281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2270..2494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2540..2968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2627..2646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2837..2858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2977..2997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2982..3311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3162..3297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3224..3243
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3378..4172
                     /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(3432..3451)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4042..4061
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4346..4467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4383..4402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4437..4456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4516..4532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4529..4551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4540..4556
                     /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(4564..4570,4571..4588,4589..4594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4609..4630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4747..4764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4918..5506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4998..5017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5677..6468,6469..6537))
                     /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     6300..6319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6538..6642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaccatgg ccagcccaag
      961 aaccaggaag gttcttaaag aagtcagggt gcaggatgag aacaacgttt gttttgagtg
     1021 tggcgcgttc aatcctcagt gggtcagtgt gacctacggc atctggatct gcctggagtg
     1081 ctcggggaga caccgcgggc ttggggttca cctcagcttt gtgcgctctg ttactatgga
     1141 caagtggaag gacattgagc ttgagaagat gaaagctggt gggaatgcta agttccgaga
     1201 gttcctggag tctcaggagg attacgatcc ttgctggtcc ttgcaggaga agtacaacag
     1261 cagagccgcg gccctcttta gggataaggt ggtcgctctg gccgaaggca gagagtggtc
     1321 tctggagtca tcacctgccc agaactggac cccacctcag cccaggacgc tgccgtccat
     1381 ggtgcaccga gtctctggcc agccgcagag tgtgaccgcc tcctcggaca aggcttttga
     1441 agactggctg aatgatgacc tcggctccta tcaaggggcc caggggaatc gctacgtggg
     1501 gtttgggaac acgccaccgc ctcagaagaa agaagatgac ttcctcaaca acgccatgtc
     1561 ctccctgtac tcgggctgga gcagcttcac cactggagcc agccggtttg cctcggcagc
     1621 caaggagggc gctacaaagt ttggatccca agcgagtcag aaggcgtccg agctgggcca
     1681 cagcctgaac gagaacgtcc tcaagcctgc gcaggagaag gtgaaggagg gaaagatttt
     1741 tgatgatgtc tccagtgggg tctctcagtt ggcgtccaag gtccagggag tcggtagtaa
     1801 gggatggcgg gacgtcacca cctttttttc ggggaaagca gagggcccct tggacagccc
     1861 ctcggagggc cacagttatc agaacagcgg tctggaccac ttccaaaaca gcaacataga
     1921 ccagagcttc tgggagacct ttggaagtgc tgagcccacc aagacccgca agtccccgag
     1981 cagcgacagc tggacgtgcg cggacacctc caccgagagg aggagctcgg acagctggga
     2041 ggtgtggggc tcggcctcca ccaacaggaa cagcaacagc gacggcgggg agggcgggga
     2101 gggcaccaag aaggcagtgc cgccggccgt gcccactgat gatggctggg acaaccagaa
     2161 ctggctcgag tatccatacg atgtaccaga ctacgcagag atccactagt tctagagggc
     2221 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2281 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2341 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2401 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2461 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2521 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2581 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2641 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2701 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2761 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2821 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2881 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2941 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3181 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3241 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3301 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3361 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3421 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3481 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3541 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3601 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3661 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3721 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3781 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3841 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3901 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3961 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4021 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4081 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4141 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4201 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4261 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4321 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4381 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4441 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6661 gccacctgac gtc
//
