LOCUS       sequence-194025-        5648 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses MAP1LC3A-V1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Nassar M, Samaha H, Ghabriel M, Yehia M, Taha H, Salem S, Shaaban K,
            Omar M, Ahmed N, El-Naggar S
  TITLE     LC3A Silencing Hinders Aggresome Vimentin Cage Clearance in Primary 
            Choroid Plexus Carcinoma.
  JOURNAL   Sci Rep. 2017 Aug 14;7(1):8022. doi: 10.1038/s41598-017-07403-5.
  PUBMED    28808307
REFERENCE   2  (bases 1 to 5648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2017-08-14"; volume: "7(1):8022. doi"; pages: " 
            10.1038/s41598-017-07403-5"
COMMENT     Sequence Label: MAP1LC3A-V1
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             635..997
                     /codon_start=1
                     /gene="MAP1LC3A"
                     /product="human LC3 protein"
                     /label=LC3A
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKD
                     EDGFLYMVYASQETFGF"
     RBS             685..693
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    join(1008..1582,1583..1585,1586..1594)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1175..1192)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1402..1421
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1595..2314
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2436..2557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2473..2492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2527..2546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2564..3019)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2701..2722)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2913..2932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3046..3150
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3152..3509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3152..3343
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3175..3195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3360..3495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3544..4338
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3544..4338
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3598..3617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4208..4227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4526..4545)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4570..4617
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4946..5534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5435..5454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgccc tcagaccggc ctttcaagca
      661 gcggcggagc ttcgccgacc gctgtaagga ggtacagcag atccgcgacc agcaccccag
      721 caaaatcccg gtgatcatcg agcgctacaa gggtgagaag cagctgcccg tcctggacaa
      781 gaccaagttt ttggtcccgg accatgtcaa catgagcgag ttggtcaaga tcatccggcg
      841 ccgcctgcag ctgaacccca cgcaggcctt cttcctgctg gtgaaccagc acagcatggt
      901 gagtgtgtcc acgcccatcg cggacatcta cgagcaggag aaagacgagg acggcttcct
      961 ctatatggtc tacgcctccc aggaaacctt cggcttctga ggatccgccc ctctccctcc
     1021 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1081 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1141 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1201 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1261 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1321 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1381 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     1441 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     1501 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     1561 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg ccgagctgtt
     1621 caccggcatc gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc acaagttcag
     1681 cgtgagcggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1741 caccaccggc aagctgcctg tgccctggcc caccctggtg accaccctga gctacggcgt
     1801 gcagtgcttc tcacgctacc ccgatcacat gaagcagcac gacttcttca agagcgccat
     1861 gcctgagggc tacatccagg agcgcaccat cttcttcgag gatgacggca actacaagtc
     1921 gcgcgccgag gtgaagttcg agggcgatac cctggtgaat cgcatcgagc tgaccggcac
     1981 cgatttcaag gaggatggca acatcctggg caataagatg gagtacaact acaacgccca
     2041 caatgtgtac atcatgaccg acaaggccaa gaatggcatc aaggtgaact tcaagatccg
     2101 ccacaacatc gaggatggca gcgtgcagct ggccgaccac taccagcaga atacccccat
     2161 cggcgatggc cctgtgctgc tgcccgataa ccactacctg tccacccaga gcgccctgtc
     2221 caaggacccc aacgagaagc gcgatcacat gatctacttc ggcttcgtga ccgccgccgc
     2281 catcacccac ggcatggatg agctgtacaa gtgagcggcc gcgactctag atcataatca
     2341 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2401 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2461 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2521 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2581 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2641 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2701 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2761 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2821 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2881 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2941 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3001 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3061 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3121 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3181 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3241 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3361 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3421 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3481 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3541 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3601 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3661 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3721 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3781 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3841 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3901 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3961 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4021 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4081 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4141 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4201 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4261 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4321 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4381 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4441 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4501 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4561 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4621 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4681 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4741 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4801 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4861 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4921 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4981 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5041 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5101 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5161 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5221 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5281 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5341 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5401 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5461 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5521 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5581 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5641 ccatgcat
//
