LOCUS       sequence-360364-        9442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SECFP-hATG9A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9442)
  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 9442)
  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-hATG9A
FEATURES             Location/Qualifiers
     source          1..9442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    427..801
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     739..755
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(833..835,836..838,839..1552)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGILTLGMDELYK"
     primer_bind     complement(878..899)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1139..1158)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1486..1507
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4205..4771
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4221..4773
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4365..4382)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4592..4611
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4785..5384
                     /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(4785..4804)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5281..5301
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5608..6077
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6207..6224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7130..7921,7922..7990))
                     /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     7753..7772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7991..8095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8163..8181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        8283..8662
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    9021..9378
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc
      421 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct
      481 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta
      541 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg
      601 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag
      661 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc
      721 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg
      781 ggtggaccat cctctagact gccggatcta gctagttaat taaggatctg ccatggtgag
      841 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      901 aaacggccac aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      961 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1021 caccctgacc tggggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1081 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga
     1141 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1201 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1261 gtacaactac atcagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1321 ggcccacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1381 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1441 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1501 gttcgtgacc gccgccggga tcttgactct cggcatggac gagctgtaca agggtggcgg
     1561 tggctcgggc ggtggtgggt cgggtggcgg cggatctatc gatgaattga attccatggc
     1621 gcagtttgac actgaatacc agcgcctaga ggcctcctat agtgattcac ccccagggga
     1681 ggaggacctg ttggtgcacg tcgccgaggg gagcaagtca ccttggcacc atattgaaaa
     1741 ccttgacctc ttcttctctc gagtttataa tctgcaccag aagaatggct tcacatgtat
     1801 gctcatcggg gagatctttg agctcatgca gttcctcttt gtggttgcct tcactacctt
     1861 cctggtcagc tgcgtggact atgacatcct atttgccaac aagatggtga accacagtct
     1921 tcaccctact gaacccgtca aggtcactct gccagacgcc tttttgcctg ctcaagtctg
     1981 tagtgccagg attcaggaaa atggctccct tatcaccatc ctggtcattg ctggtgtctt
     2041 ctggatccac cggcttatca agttcatcta taacatttgc tgctactggg agatccactc
     2101 cttctacctg cacgctctgc gcatccctat gtctgccctt ccgtattgca cgtggcaaga
     2161 agtgcaggcc cggatcgtgc agacgcagaa ggagcaccag atctgcatcc acaaacgtga
     2221 gctgacagaa ctggacatct accaccgcat cctccgtttc cagaactaca tggtggcact
     2281 ggttaacaaa tccctcctgc ctctgcgctt ccgcctgcct ggcctcgggg aagctgtctt
     2341 cttcacccgt ggtctcaagt acaactttga gctgatcctc ttctggggac ctggctctct
     2401 gtttctcaat gaatggagcc tcaaggccga gtacaaacgt ggggggcaac ggctagagct
     2461 ggcccagcgc ctcagcaacc gcatcctgtg gattggcatc gctaacttcc tgctgtgccc
     2521 cctcatcctc atatggcaaa tcctctatgc cttcttcagc tatgctgagg tgctgaagcg
     2581 ggagccgggg gccctgggag cacgctgctg gtcactctat ggccgctgct acctccgcca
     2641 cttcaacgag ctggagcacg agctgcagtc ccgcctcaac cgtggctaca agcccgcctc
     2701 caagtacatg aattgcttct tgtcacctct tttgacactg ctggccaaga atggagcctt
     2761 cttcgctggc tccatcctgg ctgtgcttat tgccctcacc atttatgacg aagatgtgtt
     2821 ggctgtggaa catgtgctga ccaccgtcac actcctgggg gtcaccgtga ccgtgtgcag
     2881 gtcctttatc ccggaccagc acatggtgtt ctgccctgag cagctgctcc gcgtgatcct
     2941 cgctcacatc cactacatgc ctgaccactg gcagggtaat gcccaccgct cgcagacccg
     3001 ggacgagttt gcccagctct tccagtacaa ggcagtgttc attttggaag agttgctgag
     3061 ccccattgtc acacccctca tcctcatctt ctgcctgcgc ccacgggccc tggagattat
     3121 agacttcttc cgaaacttca ccgtggaggt cgttggtgtg ggagatacct gctcctttgc
     3181 tcagatggat gttcgccagc atggtcatcc ccagtggcta tctgctgggc agacagaggc
     3241 ctcagtgtac cagcaagctg aggatggaaa gacagagttg tcactcatgc actttgccat
     3301 caccaaccct ggctggcagc caccacgtga gagcacagcc ttcctaggct tcctcaagga
     3361 gcaggttcag cgggatggag cagctgctgg cctcgcccaa gggggtctgc tccctgaaaa
     3421 tgccctcttt acgtctatcc agtccttaca atctgagtct gagcccctga gccttatcgc
     3481 aaatgtggta gctggctcat cctgccgggg ccctccactg cccagagacc tgcagggctc
     3541 caggcacagg gctgaagtcg cctctgccct gcgctccttc tccccgctgc aacccgggca
     3601 ggcgcccaca ggccgggcta acagcaccat gacaggctct ggggtggatg ccaggacagc
     3661 cagctccggg agcagcgtgt gggaaggaca gctgcagagc ctggtgctgt cagaatatgc
     3721 atccacagag atgagcctgc atgccctcta tatgcaccag ctccacaagc agcaggccca
     3781 ggctgaacct gagcggcatg tatggcaccg ccgggagagt gatgagagtg gagaaagcgc
     3841 ccctgatgaa gggggagagg gcgcccgggc cccccagtct atccctcgct ctgctagcta
     3901 tccctgtgca gcaccccggc ctggagctcc tgagaccacc gccctgcatg ggggcttcca
     3961 gaggcgctac ggtggcatca cagatcctgg cacagtgccc agggttccct ctcatttctc
     4021 tcggctgcct cttggagggt gggcagaaga tgggcagtcg gcatcaaggc accctgagcc
     4081 cgtgcccgaa gagggctcgg aggatgagct accccctcag gtgcacaagg tatagtttct
     4141 agaacgaatt ctgcagtcga cggtaccgcg ggcccgggat ccctcgagcg gccgctacgt
     4201 aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4261 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4321 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4381 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     4441 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     4501 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     4561 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     4621 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     4681 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     4741 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc gagtacaagc
     4801 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     4861 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     4921 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4981 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     5041 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     5101 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5161 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     5221 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     5281 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     5341 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     5401 cgcagcgccc gaccgaaagg agcgcacgac cccatggctc gacgataaaa taaaagattt
     5461 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     5521 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     5581 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     5641 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     5701 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     5761 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     5821 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     5881 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     5941 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     6001 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     6061 tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     6121 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgcgg
     7081 ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7741 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8101 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     8161 aaaataggcg tatcacgagg ccctttcgtc ttcactcgag gagcttggcc cattgcatac
     8221 gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat taccgccatg
     8281 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     8341 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     8401 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     8461 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8521 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     8581 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     8641 attagtcatc gctattacca tgctagagaa ccatcagatg tttccagggt gccccaagga
     8701 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     8761 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     8821 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     8881 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     8941 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     9001 ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc cgattgtcta
     9061 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc
     9121 tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc tgggagacgt
     9181 cccagggact tcgggggccg tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt
     9241 tggactcttt ggtgcacccc ccttagagga gggatatgtg gttctggtag gagacgagaa
     9301 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc
     9361 cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     9421 tttgtctgaa aatatgggcc cg
//
