LOCUS       sequence-176224-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf3(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/mArf3(T31N)-HA
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(338..358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        343..672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      523..658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     585..604
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             739..1533
                     /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(793..812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1403..1422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1707..1828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1744..1763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1798..1817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1877..1893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1877..1893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1890..1912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1901..1917
                     /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(1925..1931,1932..1949,1950..1955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1970..1991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2108..2125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2359..2378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3038..3829,3830..3898))
                     /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     3661..3680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3899..4003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4078..4097)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4269..4648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4649..4852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4803..4823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4897..4916
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4897..4915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4947..4956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5502..5528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5531..5547)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5531..5547)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5557..5575)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5558..5575)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5595..5612)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5601..5825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5871..5970,1..329)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(5958..5970,1..7))
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
       61 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
      121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      241 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      301 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
      361 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      421 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      481 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
      541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
      601 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
      661 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
      721 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      781 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      841 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      901 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
      961 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1021 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1081 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1141 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1201 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1261 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1321 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1381 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1441 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1501 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1561 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1621 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1681 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1741 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1801 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     1861 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1921 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1981 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2041 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2701 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4021 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     4081 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     4141 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     4201 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     4261 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     4321 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     4381 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     4441 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     4501 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     4561 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     4621 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     4681 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4741 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     4801 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     4861 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     4921 ccaagcttgg taccgagctc ggatccgcca ccatgggcaa tatctttgga aaccttctga
     4981 agagcctaat cggcaagaag gagatgcgca tcctgatggt tggcttggat gctgccggca
     5041 agaacaccat cctctacaaa ctgaaactcg gggagattgt caccaccatc ccgaccattg
     5101 ggtttaatgt ggagacagtg gaatataaga atatcagctt cacagtctgg gacgtaggtg
     5161 gccaggacaa gattcggccc ctctggagac actacttcca gaacacccaa ggcttgatat
     5221 ttgtggttga cagcaatgat cgggagcgag tgaacgaggc ccgggaagag ctgatgagga
     5281 tgctggcgga ggatgagctc cgggatgctg tgctccttgt gtttgcaaac aaacaggatt
     5341 tgccgaatgc tatgaatgct gctgagatca cagacaagct ggggctgcac tccctgcgtc
     5401 accgcaattg gtacattcag gccacctgtg ctaccagcgg ggacgggctg tatgaaggct
     5461 tggactggct ggccaatcag ctcaaaaaca agaagctcga gtatccatac gatgtaccag
     5521 actacgcaga gatccactag ttctagaggg ccctattcta tagtgtcacc taaatgctag
     5581 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     5641 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5701 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5761 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5821 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5941 cagcgcccta gcgcccgctc ctttcgcttt
//
