LOCUS       sequence-115993-        6813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of GFP-tagged LC3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  AUTHORS   Fung C, Lock R, Gao S, Salas E, Debnath J
  TITLE     Induction of autophagy during extracellular matrix detachment 
            promotes cell survival.
  JOURNAL   Mol Biol Cell. 2008 Mar . 19(3):797-806.
  PUBMED    18094039
REFERENCE   2  (bases 1 to 6813)
  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"; journalName: "Mol Biol 
            Cell. 2008 Mar . 19(3):797-806."
COMMENT     Sequence Label: pBABEpuro GFP-LC3
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(810..1409)
                     /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(893..913)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1390..1409
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1419..1748)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1433..1568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1733..1753
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(2084..2506)
                     /codon_start=1
                     /gene="Map1lc3b"
                     /product="rat LC3 protein"
                     /label=LC3
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
                     DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
     CDS             complement(join(2525..3235,3236..3238,3239..3241))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2525..3235,3236..3238,3239..3241))
                     /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(2567..2588)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2916..2935
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3175..3196
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3238..3247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3286..3489)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3315..3335)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(3538..3954)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3554..3570)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3586..3608)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4155..4354
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4419..4888
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4919..5023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5024..5092,5093..5884)
                     /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     complement(5242..5261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6044..6632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6533..6552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6786..6803
                     /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 atcgatgcat ggggtcgtgc gctcctttcg
      781 gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag
      841 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
      901 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
      961 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     1021 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     1081 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     1141 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     1201 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     1261 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     1321 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     1381 aggcgcaccg tgggcttgta ctcggtcatg gtaagctttt tgcaaaagcc taggcctcca
     1441 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     1501 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1561 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1621 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     1681 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     1741 tgacacacat tccacagggt cgactgcaga attcgcacca tagttataaa ccagggacat
     1801 gacgacgtac acaacccaca cacggcagca gtggggactt acacagtgta acataacatt
     1861 gggggttagc agggacagct gcatgcaggg agggtgggtg ctgtggggag ccaggtcgag
     1921 aggtggctgt gttggcttcc gtccgtgaca gaggcagctc tcactgagat tctcagacac
     1981 gagtgagtga gtgaccgggg atggtctgag tgtcacagtg ggctccatgc aggtagcagg
     2041 gagcagaggc ttgctttagt tggaagtggc tgtatgtctg tcacaagcat ggctctcttc
     2101 ctgttgctgt tgccttcaga gctgacatgt atgtaacagc cagtgctgtc ccgaacgtct
     2161 cctgggaggc atagaccatg tacaggaagc cgtcttcatc tctctcgctc tcgtacactt
     2221 cagagatggg tgtggacaca ctcaccatgc tgtgcccatt caccaggagg aagaaggctt
     2281 ggttagcatt gagctgcagg cgccttctaa ttatcttgat gagttcgctc atattcacgt
     2341 gatcaggtac aaggaacttg gtcttgtcca ggacgggcag ctgcttctca cccttgtatc
     2401 gctctataat cactgggatc ttggtggggt gctgctcccg gatgagccgg acatcttcca
     2461 ctctttgttc gaagctccgg cgctgtttga aggtcttctc ggacggtcta gatctgagtc
     2521 cggacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     2581 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt
     2641 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     2701 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     2761 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     2821 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     2881 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     2941 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     3001 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     3061 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     3121 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     3181 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     3241 tggtggcgac cggtagcgct agcggatctg acggttcact aaaccagctc tgcttatata
     3301 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     3361 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3421 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3481 ccgcatcacc atggtaatag cgatgactaa tacgtaccac cacactggga tccggccggc
     3541 gcctagagaa ggagtgaggg ctggataaag ggaggatcga ggcggggtcg aacgaggagg
     3601 ttcaaggggg agagacgggg cggatggagg aagaggaggc ggaggcttag ggtgtacaaa
     3661 gggcttgacc cagggagggg ggtcaaaagc caaggcttcc caggtcacga tgtaggggac
     3721 ctggtctggg tgtccatgcg ggccaggtga aaagaccttg atcttaacct gggtgatgag
     3781 gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc attctgcaga
     3841 gcagaaggta acccaacgtc tcttcttgac atctaccgac tggttgtgag cgatccgctc
     3901 gacatctttc cagtgaccta aggtcaaact taagggagtg gtaacagtct ggccctaatt
     3961 ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc tgcagcgctg
     4021 cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag acggaggcgg
     4081 gaactgtttt aggttctcgt ctcctaccag aaccacatat cctgacgggg tcggattcca
     4141 catcgactcc cttcctcagg tcgggccaca aaaacggccc ccaaagtccc tgggacgtct
     4201 cccagggttg cggccgggtg ttcagaactc gtcagttcca ccacgggtcc gccagataca
     4261 gagctagtta gctaactagt accgacgcag gcgcataaaa tcagtcatag acactagaca
     4321 atcggacaga cacagataag ttgctggcca gcttgcctcc cggtggtggg tcggtggtcc
     4381 ctgggcaggg gtctcccgat cccggacgag cccccaaatg aaagaccccc gctgacgggt
     4441 agtcaatcac tcagaggaga ccctcccaag gaacagcgag accacaagtc ggatgcaact
     4501 gcaagagggt ttattggata cacgggtacc cgggcgactc agtcaatcgg aggactggcg
     4561 ccccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag
     4621 aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg
     4681 gggcaccctg gaaacatctg atggttctct agaaactgct gagggctgga ccgcatctgg
     4741 ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt
     4801 ggcccatatt cagctgttcc atctgttctt ggccctgagc cggggcagga actgcttacc
     4861 acagatatcc tgtttggccc atattcaggc tgcaggtggc acttttcggg gaaatgtgcg
     4921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5221 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5401 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5881 gtaactgtca gaccaagttt actcatatat actttagatt gatttgcggc cgcaaaagga
     5941 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6001 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6061 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6121 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6181 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6241 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6301 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6361 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6421 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6481 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6541 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6601 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6661 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6721 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6781 agcgcagcga gtcagtgagc gaggaagcgg aag
//
