LOCUS       sequence-389386-        8971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA-tagged AP-4 epsilon in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8971)
  AUTHORS   Mattera R, Park SY, De Pace R, Guardia CM, Bonifacino JS
  TITLE     AP-4 mediates export of ATG9A from the trans-Golgi network to 
            promote autophagosome formation.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Dec 12;114(50):E10697-E10706. doi: 
            10.1073/pnas.1717327114. Epub 2017 Nov 27.
  PUBMED    29180427
REFERENCE   2  (bases 1 to 8971)
  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"; doi: 
            "10.1073/pnas.1717327114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-12-12- 12"; volume: "114"; issue: "50"; pages: 
            "E10697-E10706"
COMMENT     Sequence Label: pCI-neo-(HA)3-epsilon
FEATURES             Location/Qualifiers
     source          1..8971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        26..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     568..588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     614..638
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          778..910
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     893..912
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        955..973
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             997..1023
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1027..1053
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1060..1086
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             3364..3375
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     polyA_signal    4563..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4628..4647
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4870..5325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4957..4976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5167..5188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5442..5799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5442..5633
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5465..5485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5650..5785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5850..6644
                     /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(5904..5923)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6514..6533
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6708..6756
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6797..6816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6916..6938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6976..6994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7062..7166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7167..7235,7236..8027)
                     /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(7385..7404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8198..8786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8687..8706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactgcg atcgcccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      601 tctatataag cagagctcgt ttagtgaacc gtcagatcac tagaagcttt attgcggtag
      661 tttatcacag ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa
      721 gctgcagtga ctctcttaag gtagccttgc agaagttggt cgtgaggcac tgggcaggta
      781 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
      841 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
      901 ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac
      961 gactcactat aggctagcct cgaggccgcc accatgtacc cgtacgatgt tcctgactat
     1021 gcgggctatc cctatgacgt cccggactat gccatgggct acccttacga cgttccagat
     1081 tacgctgaat tcatgagcga catagtggag aagacgctga cggcgctgcc gggactcttt
     1141 ctgcagaacc agcccggtgg tgggcccgcg gccgccaagg cgtccttctc ctcgaggctg
     1201 ggcagccttg tccgcggcat cacagccctc acctccaagc acgaagaaga aaaattaatc
     1261 cagcaggaac tgagtagtct gaaagcgact gtttctgctc ctactacaac actgaaaatg
     1321 atgaaggaat gtatggtgag acttatatat tgtgaaatgc ttggatatga tgcttccttt
     1381 ggctatattc atgcaatcaa gttagcccaa caaggaaacc tcttagaaaa aagagtaggt
     1441 tatttggctg tttccttatt tctacatgaa agtcatgaat tattgcttct ccttgtgaat
     1501 acagttgtaa aggatctgca gagcactaac ctagtagaag tgtgtatggc actgactgtt
     1561 gttagccaga ttttcccctg cgaaatgatt ccagctgttc ttccattaat agaagataaa
     1621 cttcaacatt ctaaggagat tgtacgaaga aaagctgttc tggcattata caaattccat
     1681 ctcattgctc ctaatcaagt acaacatatt catattaagt ttcggaaagc actttgtgac
     1741 agagatgttg gggtcatggc tgcctccttg catatatatc ttagaatgat taaggaaaat
     1801 tcatctggat ataaagactt gactgggagt tttgtaacca ttttgaagca agtagttgga
     1861 ggaaagctcc cagtagagtt caattaccac agtgtgccag caccatggtt acaaattcag
     1921 ctcttgagaa tactgggact tctaggaaaa gatgatcaaa ggacaagtga attaatgtat
     1981 gatgttcttg atgaatcctt acgaagagct gagttaaatc acaatgtcac atatgctatt
     2041 ttgtttgaat gtgtgcatac agtctattct atttatccta aatcggaatt acttgagaag
     2101 gctgccaagt gcattggaaa atttgttctg tcacctaaaa taaatctaaa atatttagga
     2161 ctgaaggctc ttacctatgt tatccaacag gatcctactc tggctcttca acaccagatg
     2221 acaataattg aatgtttaga tcatcctgat cccattatta aaagagagac tctggaactt
     2281 ctttacagaa ttactaatgc acagaatata acagttattg tccagaaaat gcttgaatat
     2341 ttacatcaga gcaaagaaga gtatgtcatc gtcaatttgg tcggcaaaat agcagagctg
     2401 gctgagaaat atgctcctga taatgcatgg tttattcaga caatgaatgc tgtgttttca
     2461 gtaggaggag atgtaatgca tcctgatatt cccaataact ttctgagact actagcggaa
     2521 ggttttgatg atgaaacaga agatcagcaa ttaagactct atgcagttca gtcttatctc
     2581 actttactgg atatggaaaa tgtgttctat ccacagagat ttcttcaagt tatgagttgg
     2641 gtattagggg aatattccta cctcttagat aaggaaacgc cagaggaagt tatagctaag
     2701 ctctacaagt tacttatgaa tgactctgtg tcttcagaaa caaaagcctg gttaattgct
     2761 gctgtgacca aattgacatc tcaggcgcac tcttctaata cagttgagag attaatccat
     2821 gaatttacca tatctttgga tacttgtatg agacaacatg catttgaatt aaaacatttg
     2881 catgagaatg tggaacttat gaagagcttg cttccagttg acaggagttg tgaagacttg
     2941 gtggtagatg cttctttatc ttttctggat ggttttgtgg ctgaaggact cagtcagggt
     3001 gcagcgcctt acaaacctcc ccatcaacgc caggaggaaa agctttctca ggaaaaagtt
     3061 ctcaattttg aaccatatgg actctccttt tcttcatctg gcttcactgg acgacagtct
     3121 cctgctggca tttctcttgg ttcagatgta tctgggaata gtgctgagac aggactgaaa
     3181 gagacaaata gcttgaagct ggaaggtata aagaaattgt gggggaaaga aggctatctt
     3241 cccaagaagg aaagcaaaac tggtgatgaa agtggagctc tgcctgttcc tcaagagagt
     3301 ataatggaga atgtagatca agctataact aaaaaggatc aatctcaagt tcttacccaa
     3361 tctaaagagg agaaagaaaa gcagctgctg gcatcatcat tatttgttgg tctaggatca
     3421 gaaagtacaa tcaacctgct gggaaaagca gatactgtct ctcacaagtt cagaaggaaa
     3481 tcaaaagtca aagaagctaa aagtggcgaa acaaccagta ctcataatat gacctgttct
     3541 tcctttagtt ctttgtcaaa tgtggcatat gaagatgatt attattcgaa tactttgcac
     3601 gatacaggag acaaggaatt aaagaaattt tctctcactt cagaactttt ggattctgag
     3661 tcactcacag aactgccctt ggttgagaaa ttctcatatt gtagtctgtc tacaccttca
     3721 ttgtttgcta ataacaacat ggaaattttt caccctcctc aatctactgc agcctcagtt
     3781 gccaaggaaa gctctttagc ttcatccttt ttggaagaaa ctactgaata catacactca
     3841 aatgctatgg aagtctgtaa taatgaaact atatcagtgt cttcttataa aatttggaaa
     3901 gatgattgtt tattgatggt ctggtcagtc actaataaga gtggtttgga attgaaaagt
     3961 gctgacttag aaatttttcc tgcagaaaat ttcaaggtga ctgagcaacc tggatgctgt
     4021 ttgcctgtaa tggaagcaga aagcaccaaa agctttcaat atagtgtgca gatagaaaaa
     4081 ccttttacag aaggaaatct tactggtttt attagttatc atatgatgga tactcattct
     4141 gctcagctgg aattttctgt aaacttatca ctattagatt tcattagacc attaaaaatc
     4201 tcaagtgacg actttgggaa actctggtta tccttcgcaa atgatgtgaa acaaaatgta
     4261 aaaatgtcag aatctcaagc tgcacttcct tctgcactaa agactctgca acagaaacta
     4321 agactccata ttattgagat tataggcaat gaagggctat tggcctgtca gctgctccca
     4381 tccatcccct gcttactgca ttgccgagtt catgcagatg tattagccct gtggttcaga
     4441 tcctcctgtt ctactcttcc tgactattta ctgtatcaat gtcaaaaggt gatggaggga
     4501 tcctaggtcg actcgagcgg ccgcttccct ttagtgaggg ttaatgcttc gagcagacat
     4561 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4621 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4681 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     4741 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc cgataaggat cgatccgggc
     4801 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4861 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4921 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5161 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5221 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5281 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     5341 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     5401 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     5461 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5521 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5641 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5701 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5761 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca
     5821 acagtctcga acttaaggct agagccacca tgattgaaca agatggattg cacgcaggtt
     5881 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5941 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6001 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6061 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6121 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6181 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6241 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6301 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6361 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6421 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6481 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6541 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6601 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6661 cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt
     6721 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgcg
     6781 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6841 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6901 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6961 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     7021 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     7081 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7141 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7201 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7261 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7321 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     7381 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7441 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7501 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7561 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7621 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7681 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7741 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7801 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7861 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7921 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7981 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     8041 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     8101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8161 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8221 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8281 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8341 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8401 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8461 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8521 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8581 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8641 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8701 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8761 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8821 ccttttgctg gccttttgct cacatggctc gacagatctt caatattggc cattagccat
     8881 attattcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     8941 tctatatcat aatatgtaca tttatattgg c
//
