LOCUS       sequence-176223-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf3(Q71L) 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(Q71L)-HA
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             240..266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(269..285)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(269..285)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(295..313)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(296..313)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(333..350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    339..563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      609..1037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(696..715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     906..927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1046..1066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1051..1380
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1231..1366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1293..1312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1447..2241
                     /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(1501..1520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2111..2130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2415..2536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2452..2471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2506..2525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2585..2601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2585..2601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2598..2620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2609..2625
                     /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(2633..2639,2640..2657,2658..2663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2678..2699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2816..2833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2987..3575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3067..3086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3746..4537,4538..4606))
                     /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     4369..4388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4607..4711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4786..4805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4977..5356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5357..5560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5511..5531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5605..5624
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5605..5623
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5655..5664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gggaagagct gatgaggatg ctggcggagg atgagctccg ggatgctgtg ctccttgtgt
       61 ttgcaaacaa acaggatttg ccgaatgcta tgaatgctgc tgagatcaca gacaagctgg
      121 ggctgcactc cctgcgtcac cgcaattggt acattcaggc cacctgtgct accagcgggg
      181 acgggctgta tgaaggcttg gactggctgg ccaatcagct caaaaacaag aagctcgagt
      241 atccatacga tgtaccagac tacgcagaga tccactagtt ctagagggcc ctattctata
      301 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
      361 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
      421 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
      481 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
      541 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
      601 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      661 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      721 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      781 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      841 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      901 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      961 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1021 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     1081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     1141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1201 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     1261 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     1321 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1381 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     1441 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     1501 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     1561 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     1621 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     1681 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     1741 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     1801 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     1861 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     1921 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     1981 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2041 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2101 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2161 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2221 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     2281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2401 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2521 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     2581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3181 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3421 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4741 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     4801 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     4861 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     4921 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     4981 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5041 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5101 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5161 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5221 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5281 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5341 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5401 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5461 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5521 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     5581 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
     5641 ccgagctcgg atccgccacc atgggcaata tctttggaaa ccttctgaag agcctaatcg
     5701 gcaagaagga gatgcgcatc ctgatggtgg gcctggatgc tgccgggaag accaccatcc
     5761 tctacaagct gaaactcggg gagattgtca ccaccatccc gaccattggg tttaatgtgg
     5821 agacagtgga atataagaat atcagcttca cagtctggga cgtaggtggt ctagacaaga
     5881 ttcggcccct ctggagacac tacttccaga acacccaagg cttgatattt gtggttgaca
     5941 gcaatgatcg ggagcgagtg aacgaggccc
//
