LOCUS       sequence-176217-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf1(T31N) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5970)
  AUTHORS   Kondo Y, Hanai A, Nakai W, Katoh Y, Nakayama K, Shin HW
  TITLE     ARF1 and ARF3 are required for the integrity of recycling endosomes 
            and the recycling pathway.
  JOURNAL   Cell Struct Funct. 2012;37(2):141-54. Epub 2012 Sep 5.
  PUBMED    22971977
REFERENCE   2  (bases 1 to 5970)
  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"; journalName: "Cell Struct
            Funct."; date: "2012"; volume: "37(2)"; pages: "141-54. Epub 2012 
            Sep 5"
COMMENT     Sequence Label: pcDNA3/mArf1(T31N)-HA
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             109..903
                     /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(163..182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     773..792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1077..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1114..1133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1168..1187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1247..1263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1247..1263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1260..1282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1271..1287
                     /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(1295..1301,1302..1319,1320..1325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1340..1361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1478..1495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1649..2237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1729..1748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2408..3199,3200..3268))
                     /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     3031..3050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3269..3373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3448..3467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3639..4018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4019..4222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4173..4193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4267..4286
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4267..4285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4317..4326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4872..4898
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4901..4917)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4901..4917)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4927..4945)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4928..4945)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4965..4982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4971..5195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5241..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5328..5347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5538..5559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5678..5698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(5683..5970,1..42)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5863..5970,1..28)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5925..5944
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
       61 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
      301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
      661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
      961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     1081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1201 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     1261 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1321 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1381 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1441 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2341 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2401 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2521 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2701 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2761 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2821 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2881 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2941 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3001 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3061 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3241 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3301 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3361 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     3421 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3481 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     3541 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     3601 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     4081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     4201 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
     4261 cgaaattaat acgactcact atagggagac ccaagcttgg taccgagctc ggatccgcca
     4321 ccatggggaa catcttcgcc aacctcttca agggcctttt tggcaaaaaa gaaatgcgca
     4381 ttctcatggt gggcctggat gctgcaggga agaacacaat tctatacaaa ctgaagctgg
     4441 gcgaaattgt gaccaccatt cccaccattg gtttcaatgt ggagactgtt gaatacaaga
     4501 atatcagctt caccgtgtgg gatgtgggcg gccaggacaa gatccggccg ctgtggcgcc
     4561 actacttcca gaacacccaa ggcttgatct tcgtagtgga cagcaatgac agagagcgtg
     4621 tgaacgaggc ccgtgaagag ctcatgagga tgctagctga agatgagctc cgagatgctg
     4681 ttctcttggt gtttgccaac aagcaggacc tccccaatgc catgaatgcg gccgaaatca
     4741 cagacaagct ggggctgcac tctctacgcc acaggaactg gtacattcag gccacctgtg
     4801 ccaccagcgg ggacgggctc tatgaaggac tagattggct gtctaatcag ctccggaacc
     4861 agaagctcga gtatccatac gatgtaccag actacgcaga gatccactag ttctagaggg
     4921 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     4981 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5041 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5101 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5161 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5221 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5641 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5761 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5821 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5881 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5941 gaggccgcct ctgcctctga gctattccag
//
