LOCUS       sequence-176239-        5967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf5(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5967)
  AUTHORS   Hanai A, Ohgi M, Yagi C, Ueda T, Shin HW, Nakayama K
  TITLE     Class I Arfs (Arf1 and Arf3) and Arf6 are localized to the Flemming 
            body and play important roles in cytokinesis.
  JOURNAL   J Biochem. 2016 Feb;159(2):201-8. doi: 10.1093/jb/mvv088. Epub 2015 
            Sep 1.
  PUBMED    26330566
REFERENCE   2  (bases 1 to 5967)
  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: "10.1093/jb/mvv088";
            journalName: "J Biochem"; date: "2016-02"; volume: "159"; issue: 
            "2"; pages: "201-8"
COMMENT     Sequence Label: pcDNA3/hArf5(WT)-HA
FEATURES             Location/Qualifiers
     source          1..5967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..35
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(154..174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        159..488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      339..474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     401..420
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             555..1349
                     /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(609..628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1219..1238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1523..1644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1560..1579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1614..1633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1693..1709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1693..1709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1706..1728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1717..1733
                     /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(1741..1747,1748..1765,1766..1771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1786..1807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1924..1941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2095..2683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2175..2194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2854..3645,3646..3714))
                     /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     3477..3496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3715..3819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3894..3913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4085..4464
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4465..4668
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4619..4639
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4713..4732
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4713..4731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4763..4772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5315..5341
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5344..5360)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5344..5360)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5370..5388)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5371..5388)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5408..5425)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5414..5638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5684..5967,1..145)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5771..5790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
       61 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      121 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
      301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
      361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
      421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
      481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
      541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
      601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
      661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
      721 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
      781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
      841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
      901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
      961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     1021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     1081 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     1141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     1201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     1261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     1321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     1381 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     1441 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1501 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     1561 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1621 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     1681 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1741 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1801 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2521 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2701 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2761 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2821 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2881 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2941 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3001 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3061 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3121 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3181 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3241 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3301 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3361 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3421 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3481 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3541 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3601 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3661 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3721 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3781 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3841 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     3901 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     3961 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     4021 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     4081 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     4141 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4201 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4261 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4321 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4381 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4441 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4501 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4561 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4621 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
     4681 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
     4741 gcttggtacc gagctcggat ccgccaccat gggcctcacc gtgtccgcgc tcttttcgcg
     4801 gatcttcggg aagaagcaga tgcggattct catggttggc ttggatgcgg ctggcaagac
     4861 cacaatcctg tacaaactga agttggggga gattgtcacc accatcccaa ccataggctt
     4921 caatgtagaa acagtggaat ataagaacat ctgtttcaca gtctgggacg tgggaggcca
     4981 ggacaagatt cggcctctgt ggcggcacta cttccagaac actcagggcc tcatctttgt
     5041 ggtggacagt aatgaccggg agcgggtcca agaatctgct gatgaactcc agaagatgct
     5101 gcaggaggac gagctgcggg atgcagtgct gctggtattt gccaacaagc aggacatgcc
     5161 caacgccatg cccgtgagcg agctgactga caagctgggg ctacagcact tacgcagccg
     5221 cacgtggtat gtccaggcca cctgtgccac ccaaggcaca ggtctgtacg atggtctgga
     5281 ctggctgtcc cacgagctgt caaagcgcct cgagtatcca tacgatgtac cagactacgc
     5341 agagatccac tagttctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     5401 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5461 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5521 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5581 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     5641 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5941 gtttttcgcc ctttgacgtt ggagtcc
//
