LOCUS       sequence-224943-        7480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing GFP-YKT6 for mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  AUTHORS   Matsui T, Jiang P, Nakano S, Sakamaki Y, Yamamoto H, Mizushima N
  TITLE     Autophagosomal YKT6 is required for fusion with lysosomes 
            independently of syntaxin 17.
  JOURNAL   J Cell Biol. 2018 Aug 6;217(8):2633-2645. doi: 
            10.1083/jcb.201712058. Epub 2018 May 22.
  PUBMED    29789439
REFERENCE   2  (bases 1 to 7480)
  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: 
            "10.1083/jcb.201712058"; journalName: "J Cell Biol"; date: 
            "2018-08-6- 6"; volume: "217"; issue: "8"; pages: "2633-2645"
COMMENT     Sequence Label: pMRXIP-GFP-YKT6
FEATURES             Location/Qualifiers
     source          1..7480
                     /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(813..815,816..818,819..1529)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(813..815,816..818,819..1529)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(858..879)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1119..1138)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1466..1487
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2243..2809
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2259..2811
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2403..2420)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2630..2649
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2823..3422
                     /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(2823..2842)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3319..3339
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3646..4115
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4245..4262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5168..5959,5960..6028))
                     /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     5791..5810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6029..6133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6201..6219
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        6321..6700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    7059..7416
                     /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 gccggatctg ccatggtgag caagggcgag gagctgttca
      841 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      901 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      961 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1021 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1081 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1141 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1201 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1261 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1321 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1381 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1441 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1501 tcactctcgg catggacgag ctgtacaagg gtggcggtgg ctcgggcggt ggtgggtcgg
     1561 gtggcggcgg atctatcgat gaattgaatt cacgcgttcg cgagttaacg gatccctcga
     1621 gcggccgcat gaagctgtac agcctcagcg tcctctacaa aggcgaggcc aaggtggtgc
     1681 tgctcaaagc cgcatacgat gtgtcttcct tcagcttttt ccagagatcc agcgttcagg
     1741 aattcatgac cttcacgagt caactgattg tggagcgctc atcgaaaggc actagagctt
     1801 ctgtcaaaga acaagactat ctgtgccacg tctacgtccg gaatgatagt cttgcaggtg
     1861 tggtcattgc tgacaatgaa tacccatccc gggtggcctt taccttgctg gagaaggtac
     1921 tagatgaatt ctccaagcaa gtcgacagga tagactggcc agtaggatcc cctgctacaa
     1981 tccattaccc agccctggat ggtcacctca gtagatacca gaacccacga gaagctgatc
     2041 ccatgactaa agtgcaggcc gaactagatg agaccaaaat cattctgcac aacaccatgg
     2101 agtctctgtt agagcgaggt gagaagctag atgacttggt gtccaaatcc gaggtgctgg
     2161 gaacacagtc taaagccttc tataaaactg cccggaaaca aaactcatgc tgtgccatca
     2221 tgtgagcggc cgctacgtaa attccgcccc cccccctaac gttactggcc gaagccgctt
     2281 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     2341 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     2401 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     2461 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     2521 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     2581 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     2641 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     2701 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     2761 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     2821 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     2881 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     2941 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3001 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3061 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3121 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     3181 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     3241 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     3301 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3361 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     3421 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctcga
     3481 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     3541 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     3601 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     3661 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     3721 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     3781 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     3841 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     3901 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     3961 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4021 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4081 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     4141 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     4201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5101 acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt
     5161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5761 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     5821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6121 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6181 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cactcgagga
     6241 gcttggccca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
     6301 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     6361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6661 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctagagaacc atcagatgtt
     6721 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     6781 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     6841 cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     6901 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     6961 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
     7021 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
     7081 tgtctgtccg attgtctagt gtctatgact gattttatgc gcctgcgtcg gtactagtta
     7141 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttcgg aacacccggc
     7201 cgcaaccctg ggagacgtcc cagggacttc gggggccgtt tttgtggccc gacctgagtc
     7261 caaaaatccc gatcgttttg gactctttgg tgcacccccc ttagaggagg gatatgtggt
     7321 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     7381 ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg
     7441 tctgactgtg tttctgtatt tgtctgaaaa tatgggcccg
//
