LOCUS       sequence-412479-        8117 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 8117)
  AUTHORS   Itakura E, Mizushima N
  TITLE     Characterization of autophagosome formation site by a hierarchical 
            analysis of mammalian Atg proteins.
  JOURNAL   Autophagy. 2010 Aug;6(6):764-76.
  PUBMED    20639694
REFERENCE   2  (bases 1 to 8117)
  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"; journalName: 
            "Autophagy."; date: "2010-08"; volume: "6(6)"; pages: "764-76"
COMMENT     Sequence Label: pMXs-IP GFP-WIPI-1
FEATURES             Location/Qualifiers
     source          1..8117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..555
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(147..164)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     374..393
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             567..1169
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1066..1086
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1340..1809
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1939..1956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2110..2698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2190..2209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2862..3653,3654..3722))
                     /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     3485..3504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3723..3827)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3857..4450
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4513..4870
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4929..5345
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5275..5297
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5313..5329
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5355..5729
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5667..5683
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(5761..5763,5764..5766,5767..6477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5761..5763,5764..5766,5767..6477)
                     /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(5806..5827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6067..6086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6414..6435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
       61 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
      121 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
      181 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
      241 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
      301 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
      361 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
      421 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
      481 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
      541 aacacgatga taatatggcc acaaccatgg ttaccgagta caagcccacg gtgcgcctcg
      601 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
      661 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
      721 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
      781 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
      841 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
      901 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
      961 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     1021 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     1081 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     1141 ggtgcatgac ccgcaagccc ggtgcctgag tcgacgataa aataaaagat tttatttagt
     1201 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     1261 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     1321 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     1381 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     1441 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     1501 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     1561 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     1621 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     1681 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     1741 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     1801 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     1861 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccggccgc
     2821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3841 aaagtgccac ctgcataatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     3901 cgccattttg caaggcatgg aaaaatacat aactgagaat agaaaagttc agatcaaggt
     3961 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     4021 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     4081 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     4141 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     4201 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     4261 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     4321 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     4381 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     4441 gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca
     4501 ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat
     4561 gactgatttt atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac
     4621 ccgtggtgga actgacgagt tcggaacacc cggccgcaac cctgggagac gtcccaggga
     4681 cttcgggggc cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt tttggactct
     4741 ttggtgcacc cccctaatag gagggatatg tggttctggt aggagacgag aacctaaaac
     4801 agttcccgcc tccgtctgaa tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc
     4861 ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     4921 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     4981 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     5041 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     5101 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     5161 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     5221 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     5281 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     5341 gcgcccccat atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc
     5401 ctgaccctga catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc
     5461 tctacttagt ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg
     5521 accgaccggt ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc
     5581 agactaagaa cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca
     5641 ccgccctcaa agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg
     5701 accccggggg tggaccatcc tctagactgc cggatctagc tagttaatta aggatctgcc
     5761 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     5821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     5881 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     5941 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     6181 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     6241 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     6301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     6361 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     6421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt
     6481 ggcggtggct cgggcggtgg tgggtcgggt ggcggcggat ctatcgatga attgaattcc
     6541 gccatggagg ccgaggccgc ggacgctccc ccgggcgggg ttgagtcggc gctcagctgc
     6601 ttctctttca accaggactg cacatcccta gcaactggaa ctaaagccgg gtataagctg
     6661 ttttctctga gttctgtgga gcagctggat caagtccacg gaagcaatga aatcccggac
     6721 gtctacatcg tggagcgcct cttctccagc agcctggtgg tggtagtcag tcacacaaaa
     6781 ccacggcaga tgaacgtgta tcacttcaag aaaggcacag agatctgtaa ttacagctac
     6841 tccagcaaca tcttgtccat aaggctgaac cggcaaaggc tgctggtttg cctagaagag
     6901 tccatttata ttcacaacat taaagacatg aagctgttga agaccctcct ggatattcct
     6961 gcaaacccaa caggtctatg tgctctctct atcaaccatt ccaattctta cctggcctat
     7021 cctggaagcc tgacttcagg ggagattgtg ctttatgatg gaaactccct gaaaacagtc
     7081 tgcactattg ctgcccatga gggaacacta gctgccatca ccttcaatgc ctcaggctcc
     7141 aaactagcaa gtgcgtctga aaaaggcaca gtcatccggg tgttctctgt ccctgatggg
     7201 caaaagctct atgagttccg gagagggatg aaaaggtatg tgacaatcag ctctctagtg
     7261 ttcagtatgg attcacaatt cctctgcgcc tccagtaaca ccgagacggt acacatcttc
     7321 aagctggaac aggtcaccaa cagtcgacca gaagagcctt cgacctggag tggctacatg
     7381 ggaaagatgt ttatggctgc taccaactac ctccctaccc aggtgtcaga catgatgcat
     7441 caggacaggg cttttgccac tgcacgcttg aacttctccg gacagaggaa catctgtacc
     7501 ctctcaacga tccagaagtt gccacggctg ctagttgcgt catccagtgg acacctttat
     7561 atgtacaatt tggatcctca ggatggagga gagtgtgtct taatcaaaac ccacagcttg
     7621 cttggctcag gaacaacaga agagaataaa gaaaatgacc tcagaccttc cttacctcag
     7681 tcttatgcag cgaccgtagc cagaccaagt gcatcttcag cctccacggt gccaggttat
     7741 tctgaggacg gcggggcgct gcgaggagaa gttattcctg aacatgagtt tgcgacggga
     7801 ccagtgtgtc ttgatgatga gaatgagttt cctcctataa tcttgtgccg tggaaatcag
     7861 aagggcaaaa cgaagcagtc atgagaattc acgcgttcgc gagttaacgg atccctcgag
     7921 cggccgctac gtaaattccg cccccccccc ccctctccct cccccccccc taacgttact
     7981 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     8041 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     8101 cctassksyc yyycccc
//
