LOCUS       sequence-176242-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf5(T31N) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  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 5949)
  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(T31N)-HA
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(152..943,944..1012))
                     /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     775..794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1013..1117)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1192..1211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1383..1762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1763..1966
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1917..1937
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2011..2030
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2011..2029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2043..2052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2595..2621
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2624..2640)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2624..2640)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2650..2668)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2651..2668)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2688..2705)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2694..2918
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2964..3392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3051..3070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3261..3282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3401..3421)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3406..3735
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3586..3721
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3648..3667
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3802..4596
                     /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(3856..3875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4466..4485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4770..4891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4807..4826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4861..4880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4940..4956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4940..4956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4953..4975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4964..4980
                     /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(4988..4994,4995..5012,5013..5018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5033..5054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5171..5188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5342..5930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5422..5441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
       61 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
      181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      301 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      361 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      421 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      481 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      541 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      601 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      661 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      721 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      781 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      841 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      901 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      961 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1141 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     1201 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     1261 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     1321 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     1381 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1441 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1501 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1561 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1621 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1681 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1741 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1801 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1861 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1921 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
     1981 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     2041 ttgccaccat gggcctcacg gtgtccgcgc tcttttcgcg gatcttcggg aagaagcaga
     2101 tgcggatcct tatggttggc ctggatgcgg ccggcaagaa caccatcctc tacaagctga
     2161 agttggggga gattgtcacc accatcccca ctataggctt caatgtggaa acagtggaat
     2221 ataagaacat ctgtttcaca gtgtgggatg ttggaggcca ggataagatt cggcctctgt
     2281 ggcggcacta cttccagaac actcagggcc tcatctttgt ggtagacagc aacgaccggg
     2341 agcgggtcca ggagtctgct gatgaactcc agaagatgct gcaggaggat gagctccggg
     2401 atgcggtgct gctggtgttt gccaacaagc aggacatgcc caatgccatg cccgtgagcg
     2461 agctgactga caagctgggc cttcagcacc tgcgtagccg cacgtggtac gtccaggcca
     2521 cctgtgccac ccaaggcaca ggcctgtatg atgggctgga ctggctgtcc cacgaacttt
     2581 caaaacgtct cgagtatcca tacgatgtac cagactacgc agagatccac tagttctaga
     2641 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     2701 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2761 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2821 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2881 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     2941 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3001 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3061 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3121 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3181 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3241 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3301 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3361 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3421 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3481 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3541 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3601 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3661 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3721 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3781 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3841 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3901 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3961 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4021 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4081 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4141 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4201 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4261 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4321 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4381 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4441 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4501 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4561 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4621 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4681 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4741 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4861 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4921 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4981 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5041 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5101 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5161 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5521 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5761 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5941 tgatctttt
//
