LOCUS       sequence-252410-        7165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a wild type human Atg7/Apg7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7165)
  AUTHORS   Tanida I, Tanida-Miyake E, Ueno T, Kominami E
  TITLE     The human homolog of Saccharomyces cerevisiae Apg7p is a 
            Protein-activating enzyme for multiple substrates including human 
            Apg12p, GATE-16, GABARAP, and MAP-LC3.
  JOURNAL   J Biol Chem. 2001 Jan 19;276(3):1701-6. Epub 2000 Nov 28.
  PUBMED    11096062
REFERENCE   2  (bases 1 to 7165)
  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: "J Biol 
            Chem."; date: "2001-01-19"; volume: "276(3)"; pages: "1701-6. Epub 
            2000 Nov 28"
COMMENT     Sequence Label: pCMV-hATG7wt
FEATURES             Location/Qualifiers
     source          1..7165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3045..3054
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3051..3053,3054..3056,3057..3770)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3051..3053,3054..3056,3057..3767)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3096..3117)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3357..3376)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3704..3725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3893..4014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3930..3949)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3984..4003
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4021..4476)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4158..4179)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4370..4389
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4503..4607
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4609..4966
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4609..4800
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4632..4652)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4817..4952
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5001..5795
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5001..5795
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5055..5074)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5665..5684
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5983..6002)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6027..6074
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6403..6991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6892..6911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtatc gataagcttg
      601 gcacgaggcg gaagttgagc ggcggcaaga aataatggcg gcagctacgg gggatcctgg
      661 actctctaaa ctgcagtttg ccccttttag tagtgccttg gatgttgggt tttggcatga
      721 gttgacccag aagaagctga acgagtatcg gctggatgaa gctcccaagg acattaaggg
      781 ttattactac aatggtgact ctgctgggct gccagctcgc ttaacattgg agttcagtgc
      841 ttttgacatg agtgctccca ccccagcccg ttgctgccca gctattggaa cactgtataa
      901 caccaacaca ctcgagtctt tcaagactgc agataagaag ctccttttgg aacaagcagc
      961 aaatgagata tgggaatcca taaaatcagg cactgctctt gaaaaccctg tactcctcaa
     1021 caagttcctc ctcttgacat ttgcagatct aaagaagtac cacttctact attggttttg
     1081 ctatcctgcc ctctgtcttc cagagagttt acctctcatt caggggccag tgggtttgga
     1141 tcaaaggttt tcactaaaac agattgaagc actagagtgt gcatatgata atctttgtca
     1201 aacagaagga gtcacagctc ttccttactt cttaatcaag tatgatgaga acatggtgct
     1261 ggtttccttg cttaaacact acagtgattt cttccaaggt caaaggacga agataacaat
     1321 tggtgtatat gatccctgta acttagccca gtaccctgga tggcctttga ggaatttttt
     1381 ggtcctagca gcccacagat ggagtagcag tttccagtct gttgaagttg tttgcttccg
     1441 tgaccgtacc atgcaggggg cgagagacgt tgcccacagc atcatcttcg aagtgaagct
     1501 tccagaaatg gcatttagcc cagattgtcc taaagcagtt ggatgggaaa agaaccagaa
     1561 aggaggcatg ggaccaagga tggtgaacct cagtgaatgt atggacccta aaaggttagc
     1621 tgagtcatca gtggatctaa atctcaaact gatgtgttgg agattggttc ctactttaga
     1681 cttggacaag gttgtgtctg tcaaatgtct gctgcttgga gccggcacct tgggttgcaa
     1741 tgtagctagg acgttgatgg gttggggcgt gagacacatc acatttgtgg acaatgccaa
     1801 gatctcctac tccaatcctg tgaggcagcc tctctatgag tttgaagatt gcctaggggg
     1861 tggtaagccc aaggctctgg cagcagcgga ccggctccag aaaatattcc ccggtgtgaa
     1921 tgccagagga ttcaacatga gcatacctat gcctgggcat ccagtgaact tctccagtgt
     1981 cactctggag caagcccgca gagatgtgga gcaactggag cagctcatcg aaagccatga
     2041 tgtcgtcttc ctattgatgg acaccaggga gagccggtgg cttcctgccg tcattgctgc
     2101 aagcaagaga aagctggtca tcaatgctgc tttgggattt gacacatttg ttgtcatgag
     2161 acatggtctg aagaaaccaa agcagcaagg agctggggac ttgtgtccaa accaccctgt
     2221 ggcatctgct gacctcctgg gctcatcgct ttttgccaac atccctggtt acaagcttgg
     2281 ctgctacttc tgcaatgatg tggtggcccc aggagattca accagagacc ggaccttgga
     2341 ccagcagtgc actgtgagtc gtccaggact ggccgtgatt gcaggagccc tggccgtgga
     2401 attgatggta tctgttttgc agcatccaga agggggctat gccattgcca gcagcagtga
     2461 cgatcggatg aatgagcctc caacctctct tgggcttgtg cctcaccaga tccggggatt
     2521 tctttcacgg tttgataatg tccttcccgt cagcctggca tttgacaaat gtacagcttg
     2581 ttcttccaaa gttcttgatc aatatgaacg agaaggattt aacttcctag ccaaggtgtt
     2641 taattcttca cattccttct tagaagactt gactggtctt acattgctgc atcaagaaac
     2701 ccaagctgct gagatctggg acatgagcga cgatgagacc atctgagatg gccccgctgt
     2761 ggggctgact tctccctggc cgcctgctga ggagctctcc atcgccagag caggactgct
     2821 gaccccaggc ctggtgattc tgggcccctc ctccataccc cgaggtctgg gattcccccc
     2881 tctgctgccc aggagtggcc agtgttcggc gttgctcggg attcaagata ccaccagttc
     2941 agagctaaat aataaccttg gccttggcct tgctattgac ctgggaaaaa aaaaaaaaaa
     3001 aaaaattcct gcggccgcca ccgcgggccc gggatccacc ggtcgccacc atggtgagca
     3061 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3121 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3181 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3241 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     3301 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3361 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3421 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3481 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3541 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3601 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3661 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3721 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     3781 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3841 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3901 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3961 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     4021 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4081 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4141 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4201 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4261 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4321 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4381 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4441 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4501 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4561 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4621 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4681 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4801 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4861 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4921 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4981 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5041 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5101 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5161 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5221 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5281 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5341 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5401 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5461 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5521 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5581 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5641 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5701 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5761 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5821 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5881 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5941 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6001 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6061 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6121 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6181 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6241 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7021 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7081 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     7141 ttcatagccc atatatggag ttccg
//
