LOCUS       sequence-322830-        7635 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7635)
  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 7635)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SNAP29
FEATURES             Location/Qualifiers
     source          1..7635
                     /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(2861..3571,3572..3574,3575..3577))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2861..3571,3572..3574,3575..3577))
                     /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(2903..2924)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3252..3271
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3511..3532
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    3589..3963
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3635..3651)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3973..4389)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3989..4005)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4021..4043)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4454..4811
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     enhancer        5170..5549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5651..5669)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5737..5841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5842..5910,5911..6702)
                     /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(6060..6079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6866..7454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7355..7374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7608..7625
                     /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 tcagagttgt cgaacttttc
     2041 tttctgtgct ttttatgttg acatctaact tgtccacttt ggttgtcagc cggtcaagaa
     2101 tgtcatcttg ctcctcaatt tctgtctgca tccccagggc tatgtccttc agacgaccca
     2161 gtcccatgga cagctcatct aggttgctgt cgatcttctg gtgataggct cgaaggtgtg
     2221 ggttctttgg gtaagcatca gtactcatgg cagaaccagc ccctctgggg acagggtctg
     2281 tatcatccag ctttctaagg tttgggtggc tggcctggta ctttgcttcc tgttctttac
     2341 ttgtacttat agcttctttc aatctgttgt tgggctggga ggtgagggtg ccattctgtt
     2401 caggtggggt ctctactggt ttggatttga agtaattgac cagcccccca aacacgctct
     2461 taatgctatt gatgtgtttc tggctgatct tcaaatcttg gtccatcttg tccaccatct
     2521 tctctgtgcg ctccaggact cctcgctgac gggcgagctc ctcggaagag gcgaccccaa
     2581 ccttctcgga ctcgtacatg agggccaggg acctgctggt gctggcggcc gtggcctcag
     2641 ccctgcggag gacctcctgc cgcaagtact gctgcctgtc cgcgggcgcg tcgggcccgt
     2701 cggggaggtc tcgggcgtcc ctccaagggg ccggccgggc gccttcgtcc tccccgtcgt
     2761 cgtcgaacgg attgtagctt ttagggtaag ctgacatgga attcaattca tcgatagatc
     2821 cgccgccacc cgacccacca ccgcccgagc caccgccacc cttgtacagc tcgtccatgc
     2881 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     2941 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     3001 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     3061 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     3121 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3181 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3241 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3301 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     3361 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     3421 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     3481 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     3541 ccaccccggt gaacagctcc tcgcccttgc tcaccatggc agatccggca gtctagagga
     3601 tggtccaccc ccggggtcgg cagccttcac gtgggcggcg tgtatccaag ctgcgatgcc
     3661 gtctactttg agggcggtgg gggtggtcag caggactgtg taaggtcctt tccagcgagg
     3721 ttctaggttc ttagtctggt gtcggcggac ccacactgtg tcgccgactc ggtaagggtg
     3781 aggtaccacc ggtcggtcca gttgttcttg gtaggctgcc gccagaggtc tccagacttc
     3841 gtgctggact aagtagagag cctgtaagtg agcttggaga gaggggctgt tagtaactct
     3901 tgtcatgtca gggtcaggga agtttacaag gggcgggggt gccccatata agatctcata
     3961 tggccatatg ggggcgccta gagaaggagt gagggctgga taaagggagg atcgaggcgg
     4021 ggtcgaacga ggaggttcaa gggggagaga cggggcggat ggaggaagag gaggcggagg
     4081 cttagggtgt acaaagggct tgacccaggg aggggggtca aaagccaagg cttcccaggt
     4141 cacgatgtag gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt
     4201 aacctgggtg atgaggtctc ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt
     4261 tggccattct gcagagcaga aggtaaccca acgtctcttc ttgacatcta ccgactggtt
     4321 gtgagcgatc cgctcgacat ctttccagtg acctaaggtc aaacttaagg gagtggtaac
     4381 agtctggccc gggcccatat tttcagacaa atacagaaac acagtcagac agagacaaca
     4441 cagaacgatg ctgcagcaga caagacgcgc ggcgcggctt cggtcccaaa ccgaaagcaa
     4501 aaattcagac ggaggcggga actgttttag gttctcgtct cctaccagaa ccacatatcc
     4561 ctcctctaag gggggtgcac caaagagtcc aaaacgatcg ggatttttgg actcaggtcg
     4621 ggccacaaaa acggcccccg aagtccctgg gacgtctccc agggttgcgg ccgggtgttc
     4681 cgaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct aactagtacc
     4741 gacgcaggcg cataaaatca gtcatagaca ctagacaatc ggacagacac agataagttg
     4801 ctggccagct tacctcccgg tggtgggtcg gtggtccctg ggcaggggtc tcccgatccc
     4861 ggacgagccc ccaaatgaaa gacccccgct gacgggtagt caatcactca gaggagaccc
     4921 tcccaaggaa cagcgagacc acaagtcgga tgcaactgca agagggttta ttggatacac
     4981 gggtacccgg gcgactcagt caatcggagg actggcgcgc cgagtgaggg gttgtgggct
     5041 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt
     5101 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg
     5161 gttctctagc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     5221 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     5281 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     5341 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     5401 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     5461 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     5521 ctattaataa ctagtcaata atcaatgtca acatggcggt aatgttggac atgagccaat
     5581 ataaatgtac atattatgat atggatacaa cgtatgcaat gggccaagct cctcgagtga
     5641 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5701 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6061 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6361 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6541 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6721 tcatatatac tttagattga tttgcggccg gccgcaaaag gatctaggtg aagatccttt
     6781 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6841 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6901 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6961 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7021 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7081 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7141 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7201 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7261 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7321 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7381 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7441 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7501 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7561 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7621 gcgaggaagc ggaag
//
