LOCUS       sequence-176216-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf1(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/mArf1(Q71L)-HA
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..86
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(205..225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        210..539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      390..525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     452..471
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             606..1400
                     /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(660..679)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1270..1289
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1574..1695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1611..1630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1665..1684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1744..1760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1744..1760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1757..1779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1768..1784
                     /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(1792..1798,1799..1816,1817..1822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1837..1858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1975..1992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2146..2734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2226..2245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2905..3696,3697..3765))
                     /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     3528..3547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3766..3870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3945..3964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4136..4515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4516..4719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4670..4690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4764..4783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4764..4782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4814..4823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5369..5395
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5398..5414)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5398..5414)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5424..5442)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5425..5442)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5462..5479)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5468..5692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5738..5970,1..196)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5825..5844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
       61 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      121 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      181 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
      481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      541 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
      601 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      661 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      721 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      781 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      841 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      901 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      961 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     1021 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     1081 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     1141 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     1201 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     1261 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     1321 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     1381 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     1441 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     1501 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1561 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1621 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1681 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     1741 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2581 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3901 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     3961 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     4021 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     4081 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     4141 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4201 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4261 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4321 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4381 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4441 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4501 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4561 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4621 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4681 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
     4741 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttggtac
     4801 cgagctcgga tccgccacca tggggaacat ctttgcaaac ctcttcaagg gcctttttgg
     4861 caaaaaagaa atgcgcattc tcatggtggg cctggatgct gcagggaaga caacaattct
     4921 atacaaactg aagctgggcg aaattgtgac caccattccc accattggtt tcaatgtgga
     4981 gactgttgaa tacaagaata tcagcttcac cgtgtgggat gtgggaggcc tggacaagat
     5041 ccggccgctg tggcgccact acttccagaa cacccaaggc ttgatcttcg tagtggacag
     5101 caatgacaga gagcgtgtga acgaggcccg tgaagagctc atgaggatgc tagctgaaga
     5161 tgagctccga gatgctgttc tcttggtgtt tgccaacaag caggacctcc ccaatgccat
     5221 gaatgcggcc gaaatcacag acaagctggg gctgcactct ctacgccaca ggaactggta
     5281 cattcaggcc acctgtgcca ccagcgggga cgggctctat gaaggactag attggctgtc
     5341 taatcagctc cggaaccaga agctcgagta tccatacgat gtaccagact acgcagagat
     5401 ccactagttc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     5461 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5521 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5581 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5641 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     5701 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5761 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5821 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5881 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5941 aaaaaacttg attagggtga tggttcacgt
//
