LOCUS       sequence-248374-        8679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SECFP-hATG16A1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8679)
  AUTHORS   Koyama-Honda I, Itakura E, Fujiwara TK, Mizushima N
  TITLE     Temporal analysis of recruitment of mammalian ATG proteins to the 
            autophagosome formation site.
  JOURNAL   Autophagy. 2013 Oct;9(10):1491-9. doi: 10.4161/auto.25529. Epub 2013
            Jul 10.
  PUBMED    23884233
REFERENCE   2  (bases 1 to 8679)
  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: 
            "10.4161/auto.25529"; journalName: "Autophagy"; date: "2013-10"; 
            volume: "9"; issue: "10"; pages: "1491-9"
COMMENT     Sequence Label: pMRX-IP/SECFP-hATG16A1
FEATURES             Location/Qualifiers
     source          1..8679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(529..1128)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(612..632)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1109..1128
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1140..1692
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1142..1708
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1302..1321)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1531..1548
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(3598..4311,4312..4314,4315..4317))
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGILTLGMDELYK"
     primer_bind     complement(3643..3664)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3992..4011
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4251..4272
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4349..4723
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(4395..4411)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(4733..5149)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4749..4765)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4781..4803)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5214..5571
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     enhancer        5930..6309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(6411..6429)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6497..6601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6602..6670,6671..7462)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6820..6839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7626..8214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8115..8134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8368..8385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             join(8515..8679,1..305)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt
       61 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg
      121 gttctctaga aactgctgag ggctggaccg catctgggga ccatctgttc ttggccctga
      181 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc
      241 tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata
      301 ttcagctgtt ccatctgttc ctgaccttga tctgaacttc tctattctca gttatgtatt
      361 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc
      421 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatcg tcgagccatg
      481 gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg
      541 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
      601 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
      661 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
      721 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
      781 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
      841 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
      901 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
      961 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccgggtcgac
     1021 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggcccgggg
     1081 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat
     1141 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt
     1201 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca
     1261 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc
     1321 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg
     1381 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt
     1441 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa
     1501 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc
     1561 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg
     1621 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc
     1681 ggccagtaac gttagggggg ggggcggaat ttacgtagcg gccgctcgag ggatcccggg
     1741 cccgcggtac cgtcgactgc agaattctca gtactgtgcc cacagcacag ctttgcatcc
     1801 tttgtccaca ctgacaacgt gcgagccaga gggcgaccac gccaccgcat tgatggatga
     1861 gctgtgctgc tttgaaagaa ccttttccac tttccctgtg agcacactcc agatatacag
     1921 agagccctca gcagagcctg ccgccacgta actgccatca gggctgaaga caactctggt
     1981 ccagtcagag ccgcacttga acccaggtgc actgaatgtc tgcttgatag catttgttcg
     2041 gagatcaata acttttagca agtcatcacg ggagcagctc aggagctcag tcctttctgg
     2101 gtttaagtcc agggcagtaa tctttcccaa cagctccatc tctcgaacta tgctctctga
     2161 gcgaatgtcc cagaaacgaa ttttcttgtc aaaatgtcca ctcattacac attgctctgt
     2221 gcagacaata tcattgcaac tggatcctgc aaacactgtc tttatgcaga ctttgctgcg
     2281 tagatcccag agtttgagag tccggtcgtg acttcctgag acaatccgcg cattgtccag
     2341 caggaactta gcagacagca ctttcccact gtgtcccgtg agtgtgtgcc gtaatcgata
     2401 atcatccaca gtccagattc ggcttgcaaa atcatttgaa gctgctaaga ggtaagatcc
     2461 agcactatca aattcaatgc ttgtaattcc tgcattactg ccagataggg aacccttgaa
     2521 ctcacatttt tctccaaata cttcccaaag cttaaccctg cggtccatgc ctccagtggc
     2581 cagtaaccgg gaacctggac tgaactgcac agcgttgact tccccatcat gtgcatcgaa
     2641 gacacacaag gcagtagctg gtaccctcac ttctttacca gaaccaggat gagtatccac
     2701 attgtcctgg gggactggga aggaagagac agagcgtctc gtggctgctc tgctgatggc
     2761 tcgcacagga gaggtctctt ctgtgtgatc agaagtttca tccacaatga cctcaatgtc
     2821 atcatcctgt tcgactggta gaggttcctt tgctgcttct gcaagctctt tctgcagccg
     2881 ggcttgccgc ctcctggagt ctttttcatt ctctgcatta agccgattgg cttcctgggc
     2941 tttctcagcc atccatctgg tgaccagctc ctggttctct tccgtagttt tcctcagttt
     3001 tccctccaag gcagtaaaag tgatctgcag ggcatcatat tcatccttca gggtctggtt
     3061 ggctctttca aggtcacaaa gcttagtgcg caggtctagg cactccgtct ccaggtcaga
     3121 gatagtctgc aaacattctg caatttttgc ttcattcatc tgcatctccc tgtccttccg
     3181 ctgcatttgg ttattcaggt caatcaccag ttgagctaac tccccacgtt tcttgtgtaa
     3241 ttcagtcagt tcctcttggt gcttaatcct cagttgggcc atttcttgta gctgattgtc
     3301 attccatgtg ccatcatgtc cgggacttat ctcgtgcctg tttggtacgt catgcttttc
     3361 agcctgtagt ttctgggcca acactgaatg aagatctgac ttttccagca atttgttata
     3421 ctgcaggatg atctcctcga acgcctgtct ctgcagccgg tcccggcgcc tcagttgctc
     3481 cgagatgtgg cgcttccagc gggggaagtc ggcggcgcgg aggcccgacg acatcgaatt
     3541 caattcatcg atagatccgc cgccacccga cccaccaccg cccgagccac cgccaccctt
     3601 gtacagctcg tccatgccga gagtcaagat cccggcggcg gtcacgaact ccagcaggac
     3661 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     3721 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     3781 gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagtgggcct tgatgccgtt
     3841 cttctgcttg tcggcggtga tatagacgtt gtggctgatg tagttgtact ccagcttgtg
     3901 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     3961 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     4021 gatggtacgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     4081 catgtggtcg gggtagcggc tgaagcactg cacgccccag gtcagggtgg tcacgagggt
     4141 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     4201 ggtggcatcg ccctcgccct cgccggacac gctgaacctg tggccgttta cgtcgccgtc
     4261 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggc
     4321 agatccttaa ttaactagct agatccggca gtctagagga tggtccaccc ccggggtcgg
     4381 cagccttcac gtgggcggcg tgtatccaag ctgcgatgcc gtctactttg agggcggtgg
     4441 gggtggtcag caggactgtg taaggtcctt tccagcgagg ttctaggttc ttagtctggt
     4501 gtcggcggac ccacactgtg tcgccgactc ggtaagggtg aggtaccacc ggtcggtcca
     4561 gttgttcttg gtaggctgcc gccagaggtc tccagacttc gtgctggact aagtagagag
     4621 cctgtaagtg agcttggaga gaggggctgt tagtaactct tgtcatgtca gggtcaggga
     4681 agtttacaag gggcgggggt gccccatata agatctcata tggccatatg ggggcgccta
     4741 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa
     4801 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct
     4861 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt
     4921 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc
     4981 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga
     5041 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat
     5101 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc gggcccatat
     5161 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcaga
     5221 caagacgcgc ggcgcggctt cggtcccaaa ccgaaagcaa aaattcagac ggaggcggga
     5281 actgttttag gttctcgtct cctaccagaa ccacatatcc ctcctctaag gggggtgcac
     5341 caaagagtcc aaaacgatcg ggatttttgg actcaggtcg ggccacaaaa acggcccccg
     5401 aagtccctgg gacgtctccc agggttgcgg ccgggtgttc cgaactcgtc agttccacca
     5461 cgggtccgcc agatacagag ctagttagct aactagtacc gacgcaggcg cataaaatca
     5521 gtcatagaca ctagacaatc ggacagacac agataagttg ctggccagct tacctcccgg
     5581 tggtgggtcg gtggtccctg ggcaggggtc tcccgatccc ggacgagccc ccaaatgaaa
     5641 gacccccgct gacgggtagt caatcactca gaggagaccc tcccaaggaa cagcgagacc
     5701 acaagtcgga tgcaactgca agagggttta ttggatacac gggtacccgg gcgactcagt
     5761 caatcggagg actggcgcgc cgagtgaggg gttgtgggct cttttattga gctcggggag
     5821 cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat aaggcacagg
     5881 gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctagc atggtaatag
     5941 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     6001 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     6061 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     6121 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     6181 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     6241 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     6301 atcaatgtca acatggcggt aatgttggac atgagccaat ataaatgtac atattatgat
     6361 atggatacaa cgtatgcaat gggccaagct cctcgagtga agacgaaagg gcctcgtgat
     6421 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6481 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6541 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6601 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6661 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6721 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6781 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6841 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6901 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6961 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7021 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7081 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7141 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7201 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7261 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7321 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7381 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7441 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7501 tttgcggccg gccgcaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8401 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcaa ctatggccat
     8461 ttaatgtaaa tacttaagaa aaaaaaccaa attaattttg atacatgctg catgtgaaag
     8521 acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac agcgagacca
     8581 caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg cgactcagtc
     8641 aatcggagga ctggcgcccc gagtgagggg ttgtgggct
//
