LOCUS       sequence-492537-        9036 bp    DNA     circular SYN 11-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9036)
  AUTHORS   Itakura E, Kishi C, Inoue K, Mizushima N
  TITLE     Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes 
            with mammalian Atg14 and UVRAG.
  JOURNAL   Mol Biol Cell. 2008 Dec . 19(12):5360-72.
  PUBMED    18843052
REFERENCE   2  (bases 1 to 9036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2008 Dec . 19(12):5360-72."
COMMENT     Sequence Label: pMXs-puro Vps34-GFP
FEATURES             Location/Qualifiers
     source          1..9036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2701..2703,2704..2706,2707..3420)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2701..2703,2704..2706,2707..3417)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2746..2767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3007..3026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3354..3375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3449..3469)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3454..3783
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3634..3769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3793..4392
                     /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(3793..3812)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4289..4309
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4619..5088
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5218..5235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5389..5977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5469..5488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6141..6932,6933..7001))
                     /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     6764..6783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7002..7106)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7136..7729
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7792..8149
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             8214..8630
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8560..8582
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8598..8614
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    8640..9014
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     8952..8968
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 atgggggaag cagagaagtt tcactacatc tatagttgtg acctggatat caacgtccag
       61 cttaagatag gaagcttgga agggaagaga gaacaaaaga gttataaagc tgtcctggaa
      121 gacccaatgt tgaagttctc aggactatat caagagacat gctctgatct ttatgttact
      181 tgtcaagttt ttgcagaagg gaagcctttg gccttgccag tgagaacatc ctacaaagca
      241 tttagtacaa gatggaactg gaatgaatgg ctgaaactac cagtaaaata ccctgacctg
      301 cccaggaatg cccaagtggc cctcaccata tgggatgtgt atggtcccgg aaaagcagtg
      361 cctgtaggag gaacaacggt ttcgctcttt ggaaaatacg gcatgtttcg ccaagggatg
      421 catgacttga aagtctggcc taatgtagaa gcagatggat cagaacccac aaaaactcct
      481 ggcagaacaa gtagcactct ctcagaagat cagatgagcc gtcttgccaa gctcaccaaa
      541 gctcatcgac aaggacacat ggtgaaagta gattggctgg atagattgac atttagagaa
      601 atagaaatga taaatgagag tgaaaaacga agttctaatt tcatgtacct gatggttgaa
      661 tttcgatgtg tcaagtgtga tgataaggaa tatggtattg tttattatga aaaggacggt
      721 gatgaatcat ctccaatttt aacaagtttt gaattagtga aagttcctga cccccagatg
      781 tctatggaga atttagttga gagcaaacac cacaagcttg cccggagttt aagaagtgga
      841 ccttctgacc acgatctgaa acccaatgct gccacgagag atcagttaaa tattattgtg
      901 agttatccac caaccaagca acttacatat gaagaacaag atcttgtttg gaagtttaga
      961 tattatctta cgaatcaaga aaaagccttg acaaaattct tgaaatgtgt taattgggat
     1021 ctacctcaag aggccaaaca ggccttggaa cttctgggaa aatggaagcc gatggatgta
     1081 gaggactcct tggagctgtt atcctctcat tacaccaacc caactgtgag gcgttatgct
     1141 gttgcccggt tgcgacaggc cgatgatgag gatttgttga tgtacctatt acaattggtc
     1201 caggctctca aatatgaaaa ttttgatgat ataaagaatg gattggaacc taccaagaag
     1261 gatagtcaga gttcagtgtc agaaaatgtg tcaaattctg gaataaattc tgcagaaata
     1321 gatagctccc aaattataac cagccccctt ccttcagtct cttcacctcc tcctgcatca
     1381 aaaacaaaag aagttccaga tggcgaaaat ctggaacaag atctctgtac cttcttgata
     1441 tcgagagcct gcaaaaactc aacactggct aattatttat actggtatgt gatagtggaa
     1501 tgtgaagatc aagatactca gcagagagat ccaaagaccc atgagatgta cttgaacgta
     1561 atgagaagat tcagccaagc attgttgaag ggtgataagt ctgtcagagt tatgcgttct
     1621 ttgctggctg cacaacagac atttgtagat cggttggtgc atctaatgaa ggcagtacaa
     1681 cgcgaaagtg gaaatcgtaa gaaaaagaat gagagactac aggcattgct tggagataat
     1741 gaaaagatga atttgtcaga tgtggaactt atcccgttgc ctttagaacc ccaagtgaaa
     1801 attagaggaa taattccgga aacagctaca ctgtttaaaa gtgcccttat gcctgcacag
     1861 ttgtttttta agacggaaga tggaggcaaa tatccagtta tatttaagca tggagatgat
     1921 ttacgtcaag atcaacttat tcttcaaatc atttcactca tggacaagct gttacggaaa
     1981 gaaaatctgg acttgaaatt gacaccttat aaggtgttag ccaccagtac aaaacatggc
     2041 ttcatgcagt ttatccagtc agttcctgtg gctgaagttc ttgatacaga gggaagcatt
     2101 cagaactttt ttagaaaata tgcaccaagt gagaatgggc caaatgggat tagtgctgag
     2161 gtcatggaca cttacgttaa aagctgtgct ggatattgcg tgatcaccta tatacttgga
     2221 gttggagaca ggcacctgga taaccttttg ctaacaaaaa caggcaaact cttccacata
     2281 gactttggat atattttggg tcgggatcca aagcctcttc ctccaccaat gaagctgaat
     2341 aaagaaatgg tagaaggaat ggggggcaca cagagtgagc agtaccaaga gttccgtaaa
     2401 cagtgttaca cggctttcct ccacctgcga aggtattcta atctgatttt gaacttgttt
     2461 tccttgatgg ttgatgcaaa cattccagat attgcacttg aaccagataa aactgtgaaa
     2521 aaggttcagg ataaattccg cttagacctg tcggatgaag aggctgtgca ttacatgcag
     2581 agtctgattg atgagagtgt ccatgctctt tttgctgcag tggtggaaca gattcacaag
     2641 tttgcccagt actggagaaa agaattcact agtgttaacg gatccctcga gcggccgccc
     2701 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2761 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2821 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2881 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2941 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3001 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3061 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3121 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3181 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3241 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3301 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3361 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3421 gcggccgcca gcacagtggt cgacctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3541 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3601 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3661 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3721 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3781 aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3841 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3901 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3961 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4021 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4081 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4141 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4201 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4261 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4321 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4381 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     4441 catgcatcgt cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     4501 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     4561 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
     4621 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     4681 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     4741 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     4801 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     4861 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     4921 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     4981 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     5041 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
     5101 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     5161 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5221 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5281 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5341 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5401 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5461 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5521 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5581 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5641 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5701 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5761 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5821 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5881 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6061 aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat
     6121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga
     7141 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     7201 aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg
     7261 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     7321 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     7381 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     7441 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     7501 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     7561 agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac
     7621 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     7681 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     7741 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
     7801 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     7861 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     7921 cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg
     7981 cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc cccttagagg
     8041 agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     8101 ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc
     8161 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaatatgggc ccgggccaga
     8221 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     8281 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     8341 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     8401 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     8461 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     8521 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     8581 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc cccatatggc
     8641 catatgagat cttatatggg gcacccccgc cccttgtaaa cttccctgac cctgacatga
     8701 caagagttac taacagcccc tctctccaag ctcacttaca ggctctctac ttagtccagc
     8761 acgaagtctg gagacctctg gcggcagcct accaagaaca actggaccga ccggtggtac
     8821 ctcaccctta ccgagtcggc gacacagtgt gggtccgccg acaccagact aagaacctag
     8881 aacctcgctg gaaaggacct tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag
     8941 acggcatcgc agcttggata cacgccgccc acgtgaaggc tgccgacccc gggggtggac
     9001 catcctctag actgccggat catcgatgaa ttcgcc
//
