LOCUS       sequence-387519-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA-tagged AP-3 u3A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  AUTHORS   Mattera R, Farias GG, Mardones GA, Bonifacino JS
  TITLE     Co-assembly of viral envelope glycoproteins regulates their 
            polarized sorting in neurons.
  JOURNAL   PLoS Pathog. 2014 May 15;10(5):e1004107. doi: 
            10.1371/journal.ppat.1004107. eCollection 2014 May.
  PUBMED    24831812
REFERENCE   2  (bases 1 to 6827)
  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: "PLoS 
            Pathog."; date: "2014-05-15"; volume: "10(5):e1004107. doi"; pages: 
            " 10.1371/journal.ppat.1004107. eCollection 2014 May"
COMMENT     Sequence Label: pCI-neo-u3A-(HA)3
FEATURES             Location/Qualifiers
     source          1..6827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2392..2418
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2425..2451
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2455..2481
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2531..2652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2542..2561)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2596..2615
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2838..3293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2925..2944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3135..3156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3410..3767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3410..3601
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3433..3453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3618..3753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3818..4612
                     /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(3872..3891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4482..4501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4676..4724
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4765..4784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4884..4906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4944..4962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5030..5134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5135..5203,5204..5995)
                     /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     complement(5353..5372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6166..6754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6655..6674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt cgccaccatg atccacagtc tatttctcat caactgttct
     1141 ggcgacatat ttctagaaaa acactggaag agcgttgtaa gccaatctgt gtgtgactat
     1201 ttctttgaag ctcaggagaa agctgctgat gttgaaaatg taccacctgt catttcaaca
     1261 cctcaccact acctcattag tatctaccgg gataagctct tctttgtgtc tgtgatacag
     1321 actgaagtgc cacctctctt tgtaattgag tttctgcatc gagttgctga cacttttcag
     1381 gactactttg gtgagtgttc agaggctgca attaaggata atgtggtcat agtgtatgag
     1441 ctcttggaag aaatgttaga caatggattc ccactggcta ccgaatctaa cattctgaaa
     1501 gaactgatta aaccaccaac aattctacgt tctgtcgtca attctattac aggcagtagt
     1561 aatgttgggg acacgctccc cactgggcag ctgtccaaca tcccatggcg tcgagcaggt
     1621 gtaaagtaca ccaacaatga agcctacttt gatgtagtcg aagagataga tgcgattata
     1681 gataaatcag gatctacagt ctttgcagaa attcaaggtg tcattgatgc ttgcattaag
     1741 ctgtctggaa tgcctgatct ctctctctct ttcatgaacc caaggcttct agatgatgtc
     1801 agcttccacc catgcatccg gttcaaacgc tgggaatctg agagagtttt gtcattcatt
     1861 cctcccgatg gaaatttccg actcatatca taccgcgtca gctcacaaaa tctagtggca
     1921 atcccagtgt atgtgaaaca taatatcagc tttaaggaaa acagctcttg tggtagattt
     1981 gatataacaa ttggaccaaa acagaatatg ggaaaaacga ttgaaggaat cacagtgact
     2041 gttcacatgc caaaagttgt gctgaatatg aacctgacac caacacaagg cagctataca
     2101 ttcgatccag tcaccaaggt actagcatgg gatgtgggga aaattactcc acaaaagctc
     2161 ccaagtctta aaggactggt aaatttacag tcaggagcac ccaagccaga agagaaccca
     2221 aacctcaaca tacagttcaa gatccagcag cttgctattt caggcttaaa agtgaaccgc
     2281 ttggacatgt atggtgagaa gtataagcca tttaaaggag tcaagtatat cacaaaggcc
     2341 ggaaagttcc aagtgaggac aggatctggt tcgggaggct ctggatcagg ttacccatac
     2401 gatgttccag attacgctct cgactatccc tatgacgtcc cggactatgc cggctaccct
     2461 tacgacgttc cagattacgc ttgagcggcc gcttcccttt agtgagggtt aatgcttcga
     2521 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2581 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2641 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2701 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg
     2761 atccgggctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2821 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2881 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2941 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3001 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3061 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3121 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3181 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3241 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     3301 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag
     3361 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     3421 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt
     3781 ctgacacaac agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca
     3841 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3901 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3961 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4021 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4081 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4141 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4201 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4261 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4321 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4381 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4441 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4501 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4561 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4621 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     4681 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     4741 gatccgcgta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4801 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4861 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4921 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4981 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5041 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5101 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5161 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5221 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5281 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5341 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5401 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5461 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5521 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5581 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5641 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5701 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5821 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6001 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6301 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6781 gttcctggcc ttttgctggc cttttgctca catggctcga cagatct
//
