LOCUS       sequence-387527-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the triple Myc-tagged mouse AP-4 sigma-4 subunit in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  AUTHORS   Mattera R, De Pace R, Bonifacino JS
  TITLE     The adaptor protein chaperone AAGAB stabilizes AP-4 complex 
            subunits.
  JOURNAL   Mol Biol Cell. 2022 Oct 1;33(12):ar109. doi: 
            10.1091/mbc.E22-05-0177. Epub 2022 Aug 17.
  PUBMED    35976721
REFERENCE   2  (bases 1 to 6018)
  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: "Mol Biol 
            Cell."; date: "2022-10-1"; volume: "33(12):ar109. doi"; pages: " 
            10.1091/mbc.E22-05-0177. Epub 2022 Aug 17"
COMMENT     Sequence Label: pCI-neo-AP-4 (Myc)3-sigma-4
FEATURES             Location/Qualifiers
     source          1..6018
                     /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"
     regulatory      1097..1106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1106..1135
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1139..1168
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1175..1204
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1722..1843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1733..1752)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1787..1806
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2029..2484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2116..2135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2326..2347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2601..2958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2601..2792
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2624..2644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2809..2944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3009..3803
                     /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(3063..3082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3673..3692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3867..3915
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3956..3975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4075..4097
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4135..4153)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4221..4325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4326..4394,4395..5186)
                     /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(4544..4563)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5357..5945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5846..5865
                     /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 ctcgaggcca ccatggagca gaagctgatc tccgaagagg acctcggcga
     1141 acagaaactc atctctgagg aagatctgat gggggaacag aagctgattt ccgaggaaga
     1201 cctggaattc ggatctggtt cgggaggctc tggatcaggt atgatcaagt ttttccttat
     1261 ggtgaacaag caagggcaga cccgactgtc taagtactac gagcatgtgg acattaataa
     1321 acgtgcgctt ctggagactg aggtcagcaa gagttgcctg tctcggtcca gcgaacaatg
     1381 ctcattcatt gaatacaagg attttaaact gatctaccgg caatatgcag ctctctttgt
     1441 tgtggttgga gttaatgaca ctgagaatga gatggctatc tatgaattta tacacaattt
     1501 tgtggaagtt ttagatgggt acttcagccg agtgagtgaa ttagatataa tgtttaattt
     1561 ggataaagtt cacatcattt tggatgagat ggtgttaaat ggctgcattg tggaaactaa
     1621 cagagccaga atacttgccc ctctgctgat tcttgacaag ctgtcggaaa gctgagcggc
     1681 cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt
     1741 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1801 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1861 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     1921 ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc
     1981 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     2041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2461 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     2521 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     2581 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     2641 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2701 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2761 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2821 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2881 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2941 ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta
     3001 gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3061 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3121 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3181 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3241 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3301 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3361 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3421 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3481 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3541 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3601 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3661 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3721 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3781 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3841 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     3901 ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa
     3961 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4021 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4081 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4141 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4261 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4321 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4381 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4441 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4501 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4561 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4621 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4681 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4741 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4801 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4861 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4921 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4981 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5041 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5101 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5161 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5221 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5281 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5341 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5401 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5461 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5521 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5581 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5641 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5701 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5761 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5821 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5881 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5941 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6001 acatggctcg acagatct
//
