LOCUS       sequence-157842-        7791 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 7791)
  AUTHORS   Itakura E, Kishi-Itakura C, Mizushima N
  TITLE     The hairpin-type tail-anchored SNARE syntaxin 17 targets to 
            autophagosomes for fusion with endosomes/lysosomes.
  JOURNAL   Cell. 2012 Dec 7;151(6):1256-69. doi: 10.1016/j.cell.2012.11.001.
  PUBMED    23217709
REFERENCE   2  (bases 1 to 7791)
  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: 
            "10.1016/j.cell.2012.11"; journalName: "Cell"; date: "2012-12-7- 7";
            volume: "151"; issue: "6"; pages: "1256-69"
COMMENT     Sequence Label: pMRXIP GFP-Stx17 WT
FEATURES             Location/Qualifiers
     source          1..7791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(813..1412)
                     /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(896..916)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1393..1412
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1424..1976
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1426..1992
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1586..1605)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1815..1832
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(3017..3727,3728..3730,3731..3733))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3017..3727,3728..3730,3731..3733))
                     /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(3059..3080)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3408..3427
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3667..3688
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    3745..4119
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3791..3807)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(4129..4545)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4145..4161)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4177..4199)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4610..4967
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     enhancer        5326..5705
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5807..5825)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5893..5997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5998..6066,6067..6858)
                     /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(6216..6235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7022..7610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7764..7781
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
      121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
      241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
      301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
      541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
      601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
      661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
      721 ttctggagac taaataaaat cttttatttt atcgtcgagc catggggtcg tgcgctcctt
      781 tcggtcgggc gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac
      841 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
      901 tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
      961 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     1021 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     1081 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     1141 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     1201 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     1261 tcttgcagct cggtgacccg ctcgatgtgg cggtccgggt cgacggtgtg gcgcgtggcg
     1321 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc gggggacgtc gtcgcgggtg
     1381 gcgaggcgca ccgtgggctt gtactcggtc atggttgtgg ccatattatc atcgtgtttt
     1441 tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga cgttttttta acctcgacta
     1501 aacacatgta aagcatgtgc accgaggccc cagatcagat cccatacaat ggggtacctt
     1561 ctgggcatcc ttcagcccct tgttgaatac gcttgaggag agccatttga ctctttccac
     1621 aactatccaa ctcacaacgt ggcactgggg ttgtgccgcc tttgcaggtg tatcttatac
     1681 acgtggcttt tggccgcaga ggcacctgtc gccaggtggg gggttccgct gcctgcaaag
     1741 ggtcgctaca gacgttgttt gtcttcaaga agcttccaga ggaactgctt ccttcacgac
     1801 attcaacaga ccttgcattc ctttggcgag aggggaaaga cccctaggaa tgctcgtcaa
     1861 gaagacaggg ccaggtttcc gggccctcac attgccaaaa gacggcaata tggtggaaaa
     1921 taacatatag acaaacgcac accggcctta ttccaagcgg cttcggccag taacgttagg
     1981 ggggggggcg gaatttacgt agcggccgct cgagggatcc ttaactgcat ttcttgtcag
     2041 tttggctggg aagatctgga cagctggaag tgagcttctc catcattttc tgtttctttc
     2101 tttgtatcaa ttttccacct gtgaagccca acaccccacc accaagtgca gctgcaattc
     2161 ctgccacttt gaagcctgca aggaggccaa taggaccccc taccattccc ccgatgagtg
     2221 cacctgccac aggcagagct gccagcttgt attttgcagc cttccctaag tttttggttc
     2281 cctcttcaac attcacagca gcactgttga catggtctgc aatgctgtca atcttctcct
     2341 gctgagaatt cactaggaga gagaagtcag tgaccagttg gctaagttca attaagtccg
     2401 cttctaaggt ttcccacgat tctgcagcat tttgatcttg aggaatttca ggtaaggcat
     2461 atatctgagt caaactctga gaactagctt cagcttcagt agtatgaaat gctccaccaa
     2521 cagtcatgga tctggtcaaa ggaggctgta gcaaagtttc ttcatcatta aattgcttct
     2581 taagttcttc tacagattcc aaatggagtt ggagaaattc tgctgttgct gctgatgctt
     2641 cttctttaac aggatctatc attctcttca gaagtactag gtcatccttt cggactttca
     2701 aacaaagttt ctcaatttct cggatattgg atcggagttg ctgaactgta cgtcctgcat
     2761 tgatatgctc ttcatgcaac ttgtcccaga ttctgcacct ttgatacttc tcaatattta
     2821 tctggtgctt tcttaacctt tccaggtctg ttgggattac tatcttaatg aatttctgga
     2881 tagctggttc aagacggcgt aatttcactt tttcttcatc ttcagacatc ggatccgtta
     2941 actcgcgaac gcgtgaattc aattcatcga tagatccgcc gccacccgac ccaccaccgc
     3001 ccgagccacc gccacccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3061 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     3121 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3181 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     3241 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     3301 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3361 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     3421 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     3481 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     3541 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     3601 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     3661 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     3721 ccttgctcac catggcagat ccggcagtct agaggatggt ccacccccgg ggtcggcagc
     3781 cttcacgtgg gcggcgtgta tccaagctgc gatgccgtct actttgaggg cggtgggggt
     3841 ggtcagcagg actgtgtaag gtcctttcca gcgaggttct aggttcttag tctggtgtcg
     3901 gcggacccac actgtgtcgc cgactcggta agggtgaggt accaccggtc ggtccagttg
     3961 ttcttggtag gctgccgcca gaggtctcca gacttcgtgc tggactaagt agagagcctg
     4021 taagtgagct tggagagagg ggctgttagt aactcttgtc atgtcagggt cagggaagtt
     4081 tacaaggggc gggggtgccc catataagat ctcatatggc catatggggg cgcctagaga
     4141 aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag gttcaagggg
     4201 gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa agggcttgac
     4261 ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga cctggtctgg
     4321 gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga ggtctcggtt
     4381 aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag agcagaaggt
     4441 aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct cgacatcttt
     4501 ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggcccgggc ccatattttc
     4561 agacaaatac agaaacacag tcagacagag acaacacaga acgatgctgc agcagacaag
     4621 acgcgcggcg cggcttcggt cccaaaccga aagcaaaaat tcagacggag gcgggaactg
     4681 ttttaggttc tcgtctccta ccagaaccac atatccctcc tctaaggggg gtgcaccaaa
     4741 gagtccaaaa cgatcgggat ttttggactc aggtcgggcc acaaaaacgg cccccgaagt
     4801 ccctgggacg tctcccaggg ttgcggccgg gtgttccgaa ctcgtcagtt ccaccacggg
     4861 tccgccagat acagagctag ttagctaact agtaccgacg caggcgcata aaatcagtca
     4921 tagacactag acaatcggac agacacagat aagttgctgg ccagcttacc tcccggtggt
     4981 gggtcggtgg tccctgggca ggggtctccc gatcccggac gagcccccaa atgaaagacc
     5041 cccgctgacg ggtagtcaat cactcagagg agaccctccc aaggaacagc gagaccacaa
     5101 gtcggatgca actgcaagag ggtttattgg atacacgggt acccgggcga ctcagtcaat
     5161 cggaggactg gcgcgccgag tgaggggttg tgggctcttt tattgagctc ggggagcaga
     5221 agcgcgcgaa cagaagcgag aagcgaactg attggttagt tcaaataagg cacagggtca
     5281 tttcaggtcc ttggggcacc ctggaaacat ctgatggttc tctagcatgg taatagcgat
     5341 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     5401 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     5461 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     5521 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     5581 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     5641 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     5701 atgtcaacat ggcggtaatg ttggacatga gccaatataa atgtacatat tatgatatgg
     5761 atacaacgta tgcaatgggc caagctcctc gagtgaagac gaaagggcct cgtgatacgc
     5821 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5881 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5941 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6001 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6061 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6121 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6181 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6241 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6301 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6361 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6421 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6481 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6541 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6601 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6661 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6721 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6781 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6841 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgatttg
     6901 cggccggccg caaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
