LOCUS       sequence-341981-        8868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RFP-hATG9A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8868)
  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 8868)
  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: pMXs-puro-RFP-ATG9A
FEATURES             Location/Qualifiers
     source          1..8868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1079..1495
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1425..1447
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1463..1479
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1505..1879
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1817..1833
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1896..2570
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1896..2570
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2026..2044)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2294..2313
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2488..2511
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5182..5202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5187..5516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5367..5502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5526..6125
                     /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(5526..5545)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6022..6042
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6352..6821
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6951..6968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7122..7710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7202..7221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7874..8665,8666..8734))
                     /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     8497..8516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8735..8839)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt
      901 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg
     1081 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1141 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1201 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1261 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1321 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1381 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1441 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1501 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1561 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1621 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1681 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     1741 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     1801 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     1861 tggaccatcc tctagactgc cggatcatcg ataccatggc ctcctccgag gacgtcatca
     1921 aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga
     1981 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2041 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc cagtacggct
     2101 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2161 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2221 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2281 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     2341 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gatgaggctg aagctgaagg
     2401 acggcggcca ctacgacgcc gaggtcaaga ccacctacat ggccaagaag cccgtgcagc
     2461 tgcccggcgc ctacaagacc gacatcaagc tggacatcac ctcccacaac gaggactaca
     2521 ccatcgtgga acagtacgag cgcgccgagg gccgccactc caccggcgcc atcgatgaat
     2581 tgaattccat ggcgcagttt gacactgaat accagcgcct agaggcctcc tatagtgatt
     2641 cacccccagg ggaggaggac ctgttggtgc acgtcgccga ggggagcaag tcaccttggc
     2701 accatattga aaaccttgac ctcttcttct ctcgagttta taatctgcac cagaagaatg
     2761 gcttcacatg tatgctcatc ggggagatct ttgagctcat gcagttcctc tttgtggttg
     2821 ccttcactac cttcctggtc agctgcgtgg actatgacat cctatttgcc aacaagatgg
     2881 tgaaccacag tcttcaccct actgaacccg tcaaggtcac tctgccagac gcctttttgc
     2941 ctgctcaagt ctgtagtgcc aggattcagg aaaatggctc ccttatcacc atcctggtca
     3001 ttgctggtgt cttctggatc caccggctta tcaagttcat ctataacatt tgctgctact
     3061 gggagatcca ctccttctac ctgcacgctc tgcgcatccc tatgtctgcc cttccgtatt
     3121 gcacgtggca agaagtgcag gcccggatcg tgcagacgca gaaggagcac cagatctgca
     3181 tccacaaacg tgagctgaca gaactggaca tctaccaccg catcctccgt ttccagaact
     3241 acatggtggc actggttaac aaatccctcc tgcctctgcg cttccgcctg cctggcctcg
     3301 gggaagctgt cttcttcacc cgtggtctca agtacaactt tgagctgatc ctcttctggg
     3361 gacctggctc tctgtttctc aatgaatgga gcctcaaggc cgagtacaaa cgtggggggc
     3421 aacggctaga gctggcccag cgcctcagca accgcatcct gtggattggc atcgctaact
     3481 tcctgctgtg ccccctcatc ctcatatggc aaatcctcta tgccttcttc agctatgctg
     3541 aggtgctgaa gcgggagccg ggggccctgg gagcacgctg ctggtcactc tatggccgct
     3601 gctacctccg ccacttcaac gagctggagc acgagctgca gtcccgcctc aaccgtggct
     3661 acaagcccgc ctccaagtac atgaattgct tcttgtcacc tcttttgaca ctgctggcca
     3721 agaatggagc cttcttcgct ggctccatcc tggctgtgct tattgccctc accatttatg
     3781 acgaagatgt gttggctgtg gaacatgtgc tgaccaccgt cacactcctg ggggtcaccg
     3841 tgaccgtgtg caggtccttt atcccggacc agcacatggt gttctgccct gagcagctgc
     3901 tccgcgtgat cctcgctcac atccactaca tgcctgacca ctggcagggt aatgcccacc
     3961 gctcgcagac ccgggacgag tttgcccagc tcttccagta caaggcagtg ttcattttgg
     4021 aagagttgct gagccccatt gtcacacccc tcatcctcat cttctgcctg cgcccacggg
     4081 ccctggagat tatagacttc ttccgaaact tcaccgtgga ggtcgttggt gtgggagata
     4141 cctgctcctt tgctcagatg gatgttcgcc agcatggtca tccccagtgg ctatctgctg
     4201 ggcagacaga ggcctcagtg taccagcaag ctgaggatgg aaagacagag ttgtcactca
     4261 tgcactttgc catcaccaac cctggctggc agccaccacg tgagagcaca gccttcctag
     4321 gcttcctcaa ggagcaggtt cagcgggatg gagcagctgc tggcctcgcc caagggggtc
     4381 tgctccctga aaatgccctc tttacgtcta tccagtcctt acaatctgag tctgagcccc
     4441 tgagccttat cgcaaatgtg gtagctggct catcctgccg gggccctcca ctgcccagag
     4501 acctgcaggg ctccaggcac agggctgaag tcgcctctgc cctgcgctcc ttctccccgc
     4561 tgcaacccgg gcaggcgccc acaggccggg ctaacagcac catgacaggc tctggggtgg
     4621 atgccaggac agccagctcc gggagcagcg tgtgggaagg acagctgcag agcctggtgc
     4681 tgtcagaata tgcatccaca gagatgagcc tgcatgccct ctatatgcac cagctccaca
     4741 agcagcaggc ccaggctgaa cctgagcggc atgtatggca ccgccgggag agtgatgaga
     4801 gtggagaaag cgcccctgat gaagggggag agggcgcccg ggccccccag tctatccctc
     4861 gctctgctag ctatccctgt gcagcacccc ggcctggagc tcctgagacc accgccctgc
     4921 atgggggctt ccagaggcgc tacggtggca tcacagatcc tggcacagtg cccagggttc
     4981 cctctcattt ctctcggctg cctcttggag ggtgggcaga agatgggcag tcggcatcaa
     5041 ggcaccctga gcccgtgccc gaagagggct cggaggatga gctaccccct caggtgcaca
     5101 aggtatagtt tctagaacga attcacgcgt tcgcgagtta acggatccct cgagcggccg
     5161 ccagcacagt ggtcgacctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5221 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5281 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5341 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5401 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5461 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5521 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5581 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5641 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5701 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5761 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5821 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5881 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5941 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     6001 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     6061 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     6121 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
     6181 cgtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     6241 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     6301 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     6361 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     6421 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     6481 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     6541 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     6601 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     6661 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     6721 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     6781 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     6841 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     6901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7801 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     7861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8161 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8461 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8821 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcat
//
