LOCUS       sequence-387522-        6842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA-tagged AP-1 u1A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6842)
  AUTHORS   Guo X, Mattera R, Ren X, Chen Y, Retamal C, Gonzalez A, Bonifacino 
            JS
  TITLE     The adaptor protein-1 mu1B subunit expands the repertoire of 
            basolateral sorting signal recognition in epithelial cells.
  JOURNAL   Dev Cell. 2013 Nov 11;27(3):353-66. doi: 
            10.1016/j.devcel.2013.10.006.
  PUBMED    24229647
REFERENCE   2  (bases 1 to 6842)
  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: 
            "10.1016/j.devcel.2013.10"; journalName: "Dev Cell"; date: 
            "2013-11-11- 11"; volume: "27"; issue: "3"; pages: "353-66"
COMMENT     Sequence Label: pCI-neo-u1A-(HA)3
FEATURES             Location/Qualifiers
     source          1..6842
                     /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      1260..1269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2407..2433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2440..2466
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2470..2496
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2546..2667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2557..2576)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2611..2630
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2853..3308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2940..2959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3150..3171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3425..3782
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3425..3616
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3448..3468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3633..3768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3833..4627
                     /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(3887..3906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4497..4516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4691..4739
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4780..4799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4899..4921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4959..4977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5045..5149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5150..5218,5219..6010)
                     /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(5368..5387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6181..6769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6670..6689
                     /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 tccgccagcg ccgtctacgt actggatctg
     1141 aagggcaagg tgctcatctg caggaactac cgtggggatg tggacatgtc agaggtggag
     1201 cacttcatgc ccattctgat ggagaaggag gaggagggca tgctgtcacc tatcttggcc
     1261 catggtggcg ttcgtttcat gtggattaag cacaacaacc tgtacctggt cgccacttca
     1321 aaaaagaatg cttgtgtgtc actggtgttc tccttcctct acaaggtggt acaggtcttc
     1381 tccgagtact ttaaggagtt ggaggaggag agcatccgag acaactttgt catcatctac
     1441 gagctgctag atgagctcat ggactttggc tacccgcaga ccactgacag caagatcttg
     1501 caggagtaca tcactcagga aggccacaag ctggaaacgg gggcccctag gcccccagcc
     1561 acagtcacca atgctgtgtc ctggcgttca gaaggcatca agtatcggaa gaatgaagta
     1621 ttcctggatg tcattgaggc tgttaacctc ttggtcagtg ccaatggcaa cgtgctgcgc
     1681 agtgagattg tgggttccat caagatgcgg gtcttcctct caggcatgcc tgagttacgc
     1741 ctgggtctca atgacaaggt cctcttcgac aacacaggcc gagggaagag caagtcagtg
     1801 gagctggagg atgtgaaatt ccaccagtgt gtgcggctgt cacgttttga gaacgaccgc
     1861 actatctcct tcatcccacc cgacggagag tttgaactca tgtcctaccg cctcaacacc
     1921 catgtgaagc ctttgatctg gattgagtcc gtgattgaga agcattccca cagccgcatt
     1981 gagtacatgg tcaaggccaa gagccagttc aagaggcggt caacagccaa caatgtagag
     2041 atccatatac cagtccccaa cgatgctgat tcacccaagt tcaagactac agtggggagt
     2101 gtcaagtggg tccctgaaaa cagtgagatc gtgtggtccg tcaagtcctt tccgggtggc
     2161 aaggagtacc tgatgcgggc ccactttggc cttcccagtg tggaagctga agacaaggag
     2221 ggaaagcccc ccatcagcgt caagtttgag atcccctact tcactacctc tggcatccag
     2281 gtgcgctacc tgaaaatcat tgagaagagt gggtaccagg ccctgccctg ggtacgatat
     2341 atcacacaga acggagatta tcagctccgg acccagggat ctggttcggg aggctctgga
     2401 tcaggttacc catacgatgt tccagattac gctgtcgact atccctatga cgtcccggac
     2461 tatgccggct acccttacga cgttccagat tacgcttgag cggccgcttc cctttagtga
     2521 gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     2581 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2641 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2701 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     2761 atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2821 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2881 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2941 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3001 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3061 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3121 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3181 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3241 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3301 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3361 cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
     3421 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3541 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3601 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3721 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3781 aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca ccatgattga
     3841 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3901 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3961 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4021 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4081 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4141 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4201 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4261 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4321 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4381 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4441 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4501 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4561 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4621 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4681 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4741 atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct ctgatgccgc
     4801 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4861 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4921 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4981 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5041 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5101 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5161 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5221 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5281 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5341 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5401 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5461 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5521 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5581 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5641 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5701 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5761 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5821 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5881 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5941 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6001 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6061 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6121 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6181 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6241 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6301 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6661 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6721 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6781 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
     6841 ct
//
