LOCUS       sequence-176222-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf3(WT) 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/hArf3(WT)-HA
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..58)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     249..270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(389..409)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        394..723
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      574..709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     636..655
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             790..1584
                     /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(844..863)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1454..1473
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1758..1879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1795..1814)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1849..1868
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1928..1944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1928..1944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1941..1963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1952..1968
                     /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(1976..1982,1983..2000,2001..2006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2021..2042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2159..2176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2330..2918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2410..2429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3089..3880,3881..3949))
                     /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     3712..3731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3950..4054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4129..4148)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4320..4699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4700..4903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4854..4874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4948..4967
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4948..4966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4998..5007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5553..5579
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5582..5598)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5582..5598)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5608..5626)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5609..5626)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5646..5663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5652..5876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5922..5970,1..380)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
       61 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      121 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      181 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      241 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      301 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      361 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      421 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      541 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      601 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      661 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      721 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
      781 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
      841 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
      901 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
      961 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     1021 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     1081 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     1141 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     1201 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     1261 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     1321 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     1381 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     1441 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     1501 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     1561 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     1621 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     1681 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     1741 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1801 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1861 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     1921 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1981 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2161 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2221 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2521 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2761 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3001 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3061 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3121 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3181 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3241 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3301 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3361 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3421 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3481 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3541 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3601 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3661 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3721 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3781 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3841 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3901 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3961 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4021 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4081 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     4141 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     4201 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     4261 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     4321 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     4381 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4441 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4501 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4561 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4621 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     4681 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     4741 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     4801 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     4861 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
     4921 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
     4981 gtaccgagct cggatccgcc accatgggca atatctttgg aaaccttctc aagagcctga
     5041 ttgggaagaa ggagatgcgc atcctgatgg tgggcctgga tgccgcagga aagaccacca
     5101 tcctatacaa gctgaaactg ggggagatcg tcaccaccat ccctaccatt gggttcaatg
     5161 tggagacagt ggagtataag aacatcagct ttacagtgtg ggatgtgggt ggccaggaca
     5221 agattcgacc cctctggaga cactacttcc agaacaccca agggttgata tttgtggtcg
     5281 acagcaatga tcgggagcga gtaaatgagg cccgggaaga gctgatgaga atgctggcgg
     5341 aggacgagct ccgggatgct gtactccttg tctttgcaaa caaacaggat ctgcctaatg
     5401 ctatgaacgc tgctgagatc acagacaagc tgggcctgca ttcccttcgt caccgtaact
     5461 ggtacattca ggccacctgt gccaccagcg gggacgggct gtacgaaggc ctggactggc
     5521 tggccaatca gctcaaaaac aagaagctcg agtatccata cgatgtacca gactacgcag
     5581 agatccacta gttctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     5641 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     5701 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     5761 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     5821 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     5881 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5941 attaagcgcg gcgggtgtgg tggttacgcg
//
