LOCUS       sequence-175308-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8078)
  AUTHORS   Mizushima N, Kuma A, Kobayashi Y, Yamamoto A, Matsubae M, Takao T, 
            Natsume T, Ohsumi Y, Yoshimori T
  TITLE     Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic 
            isolation membrane with the Apg12-Apg5 conjugate.
  JOURNAL   J Cell Sci. 2003 May 1. 116(Pt 9):1679-88.
  PUBMED    12665549
REFERENCE   2  (bases 1 to 8078)
  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 Cell Sci.
            2003 May 1. 116(Pt 9):1679-88."
COMMENT     Sequence Label: p3xFLAG-CMV10-mApg16L
FEATURES             Location/Qualifiers
     source          1..8078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(253..273)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        271..568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             622..1416
                     /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(676..695)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1286..1305
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2068..2202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2105..2124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2159..2178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2207..2226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2208..2226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2240..2256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2240..2256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2253..2275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2264..2280
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2288..2294,2295..2312,2313..2318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2333..2354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2471..2488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2642..3230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2722..2741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3401..4192,4193..4261))
                     /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     4024..4043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4262..4366)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4393..4848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4690..4711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4974..4996
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4988..5005
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4989..5005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5166..5545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5546..5749
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5700..5720
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5779..5844
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    join(7685..8078,1..229)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7884..7903)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
       61 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
      121 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
      181 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
      241 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
      301 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      361 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      421 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      481 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
      541 tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag
      601 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
      661 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
      721 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
      781 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
      841 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
      901 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
      961 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     1021 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     1081 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     1141 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     1201 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     1261 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     1321 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     1381 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     1441 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     1501 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     1561 atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct
     1621 gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc
     1681 aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca
     1741 cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc
     1801 ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga
     1861 gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg
     1921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc
     1981 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2041 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2101 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2161 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc
     2221 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2281 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2341 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2401 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2821 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3061 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3541 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3601 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3661 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3721 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3781 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3841 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3901 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3961 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4021 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4081 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4141 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4201 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4261 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4321 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4381 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4441 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4501 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4561 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4621 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4681 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4741 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4801 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     4861 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     4921 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     4981 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     5041 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     5101 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     5161 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     5221 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5281 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5341 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5401 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     5461 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     5521 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     5581 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     5641 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     5701 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     5761 ccgtcagaat taaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc
     5821 gattacaagg atgacgatga caagcttgcg gccgcgaatt catcgataga tctgatatcg
     5881 gtaccagtcg actctagatt cggcatgtcg tcgggcctgc gcgccgcaga ctttccccgc
     5941 tggaagcgtc acatcgcgga ggaactgagg cgccgggacc gactgcagag gcaggcgttc
     6001 gaggagatca ttctgcagta taccaagttg ctggaaaagt cagatcttca ttcagtattg
     6061 acccagaaac tacaagcaga aaagcatgac atgccaaata ggcatgaaat aagtcctgga
     6121 catgatggtg cgtggaatga tagtcaacta caagaaatgg cccagttgag gatcaaacac
     6181 caggaagagc tgaccgaact gcacaagaag cgtggggagt tagctcagtt ggtgattgac
     6241 ctgaacaacc aaatgcagca gaaggacaag gagatacaga tgaatgaagc aaagatttcg
     6301 gagtatttac agaccatctc tgacctggag acaaactgcc tggacctgcg caccaaactg
     6361 caggacctcg aggtagccaa ccagaccctg aaggatgagt atgacgccct gcagattact
     6421 tttactgccc tagaagagaa actgaggaaa actactgagg agaaccagga actggtcacc
     6481 agatggatgg ctgagaaggc ccaagaagcc aatcgcctca atgcagagaa tgagaaggac
     6541 tccaggaggc gtcaagcacg gctgcagaag gagcttgcag aagcagcaaa ggaacctcta
     6601 cctgttgaac aggatgatga cattgaagtc attgtggatg agacctcaga ccacacagaa
     6661 gagacctctc ccgtccgagc tgtcagcaga gcagctacga gacgctctgt ctcttccatc
     6721 ccagtccccc aggatatcat ggacactcat cctgcttctg gtaaagatgt gagagtccca
     6781 actactgcct cgtatgtctt cgatgcgcat gacggagagg tcaacgcagt gcagttcagt
     6841 ccaggctccc ggttgctggc cactggaggc atggaccgca gggtgaaact ttgggaagca
     6901 ttcggagata aatgtgaatt caagggctcc ctgtctggca gtaatgctgg aattacaagc
     6961 attgaatttg atagtgctgg agcttaccta ttagcagctt caaatgattt tgcaagccga
     7021 atctggactg tggatgatta tcgattacgg cacacactca caggccacag cgggaaagtc
     7081 ctctctgcca agttcctgct ggacaatgca cggattgtct caggaagtca cgaccggacc
     7141 ctcaaactct gggatctccg cagcaaagtc tgcataaaaa cagtgtttgc aggatccagc
     7201 tgcaatgaca ttgtttgcac tgaacaatgt gtaatgagtg gacattttga caagaaaatt
     7261 cgtttctggg atatccggtc agagagtgtg gtccgagaga tggaactgtt agggaagatc
     7321 actgctctgg acctaaaccc tgagagaact gagctcctga gctgctcccg tgatgacctg
     7381 ctaaaagtca tcgacctccg gacaaatgca gtcaaacaga cattcagtgc gcctggattc
     7441 aaatgcggct ctgactggac ccgggttgtc ttcagccctg atggcagtta cgtggcagca
     7501 ggctcagccg agggttctct ttatgtctgg agtgtgctga cagggaaagt ggagaaggtt
     7561 ctttcaaaac agcacagctc ttctatcaat gcggtggcgt gggccccctc gggcttacat
     7621 gttgtcagtg tggacaaagg aagcagagct atgctgtggg cacagccttg atctagagga
     7681 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     7741 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     7801 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     7861 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     7921 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     7981 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     8041 acggggtttc accatattgg ccaggctggt ctccaact
//
