LOCUS       sequence-122825-       10731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lgg-1 tagged with oxCerulean-oxVenus (dFP, or double fluorescent 
            protein), expressed in intestines, for monitoring autophagy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10731)
  AUTHORS   Chapin HA, Okada M, Merz AJ, Miller DL
  TITLE     Tissue-specific autophagy responses to aging and stress in C. 
            elegans.
  JOURNAL   Aging (Albany NY). 2015 Jun 30.
  PUBMED    26142908
REFERENCE   2  (bases 1 to 10731)
  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: "Aging 
            (Albany NY). 2015 Jun 30."
COMMENT     Sequence Label: pVha-6::oxCerulean-oxVenus::Lgg-1
FEATURES             Location/Qualifiers
     source          1..10731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(214..233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     333..355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(393..411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        479..583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(584..652,653..1444)
                     /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(802..821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1615..2203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2104..2123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3839..3850)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    5916..5936
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    6818..6842
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7673..7693
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(8445..8469)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9199..9219)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(9227..9244)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9227..9243)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cacgtgacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
       61 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      121 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
      181 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      241 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
      361 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
      421 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
      481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      601 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      661 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      721 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      781 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      841 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      901 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      961 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1021 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1081 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1141 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1201 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1261 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1321 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1381 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1441 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1501 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1561 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1621 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1681 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1741 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     1801 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1861 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1921 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1981 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2041 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2101 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2161 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2221 ctttttacgg ttcctggtct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2281 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgatacca gcggataaca
     2341 atttcacagg gccctgtcgg attatgggct cttcttttat cctctacttt ctttctagaa
     2401 aatgcaaatg ttgagtcaga aattgctgaa tggagtacaa ctgcaatggg tttagcttat
     2461 gtttctggaa caattactgg tggaattgtg attgaaaagt aaggttatat ttcatttcaa
     2521 taaatgttac cacagtataa caaaaacaag ttacattgtt cagagtatgt ctgaatgtat
     2581 tattattatt gtatttgtgc atcagtactg tagttgtact gaagaagggt ttaaaataat
     2641 ctcaacgttt ttaagacgag ttagtgagaa tacggtcgca gttaggatgt ttggacatag
     2701 ttttattaga taatcgtgaa ctaactcact ataaaaaccc gctaactcat tgaataagat
     2761 agttttaaac gttcaaaatt tgaagacaaa attgtatgaa taagtttaca gagttggtcg
     2821 acgcaagctt ctccttctct tcacatttct caataattta gctctaattg cattcgtatt
     2881 cttcgcaaaa atacgtattc tgattgaccc gatgaaatac ggatgcctag ttgcattgat
     2941 catctacggt tatacttatg ggtgaggttt ttcaaatata ttttgattta atcgaataat
     3001 gaaatttcag aactggagtt ggtccaattt cgtggtttat atcctctgaa ctggtacctc
     3061 aaaaacatcg tagtattgct caatctgtgg catattcaat gaatactgta atggtggtta
     3121 tcagtacatt tacagtatta cctctatatt ccgttattgg tagttatgct ttccttattc
     3181 tatattcaat tccttcattt ataagtatgc ttatattatt tcgctactta ccggaaacca
     3241 aaggacgaga gattcatgat attgtgaatg aacttaaaaa aaaatgaaaa cctttgaaaa
     3301 cgttcatacg gccgaaattt tacttgatcg agccaattga acttttgtta agacttacag
     3361 aaaagtgaga gattttgtta caaaaaagtg agaaaaaagc gaaaatgaac aggaaaaaga
     3421 agaaaaatac acccgaagta tcatataaaa attaaaaaga ttgtttgacc tggcggaact
     3481 tttttgcaaa aactctgaat atccatggca catgcttcgt gcaaaacagg atccaaatga
     3541 atatcaacaa gagattcttg aagacgacga gccacttgct cagagcattg cttatctcga
     3601 atgagaccct tgttaaaatc atttttcaaa cattatatga cattgtcgaa atgtcctcct
     3661 gattccatga tggtagcaaa ctcacttcgt ctttgagcca atttatccaa gtccttgtaa
     3721 aaagtattcg aaaattgtta acggataaaa tgtttattat aatatcaaaa acaatttgtc
     3781 agttgaccac tttttgatat atttgacaga aaccgggatg aattggctca aagttagggc
     3841 gcccttctat tacagggttt ctgataacaa accggttatt aactcccaac aagggatgat
     3901 ttcaattcat catgctcaaa ttgacccaaa ttaagttaca tgacaaattc atcgcccttt
     3961 tcaactcttc ttggctcatc atctgttatt ctgttcctat tctgtctgca ccctataccc
     4021 ctttgcatac tctctcgtca ttcctttctt ggatagtgtc ttcttctccc cagctctgct
     4081 acttctatga cttgcgcccg ctgcttttcc gctcgttcct ctcctctctg acgtcttctt
     4141 ctgcttcttt tctccttcct ttttcccttc tcaatcttcc tctttttccc atttctgtca
     4201 atcattcgaa gaagaagaag aagaagaccc tcattcattt catttttttt tctgtcggtg
     4261 tgtgctgctc ggttaagagt gagctctcta ttccacgtct tcttcttttt cttcttcgat
     4321 tcgaatcaat cactccacca aaacgcattc gtttttggga ttcaccccgc ggttcgcaac
     4381 aggtttcttt ttcaaattat tagcgttata aatagaaaaa tggtggagtt tcaataaaaa
     4441 tgataattta caaagtgtat tttgattaca tgtactcaaa aaggttgaaa tttttcatac
     4501 cagttttccg gaaatccatc tgatatcatt atcgtatttt tctttttaaa aatgttttca
     4561 aaaaaaaaca aaatatagct ggttatttgg caccctctaa ttaccatttt cctgtcacac
     4621 cacactcttt tccttcttcc acttctttcc gttttcagcc gcttccaacc aaaccgatat
     4681 gaaagccgag caacaacaat cgattccacc cggctcggcg accttcccgt cgcaggtgag
     4741 actcagaaaa ctagagaaac cgctcaacta actcttgata tccgatttca ttcttttctc
     4801 ttttcttttt gttgaacttt tccatatttc cagatgccac ggccaccacc aagcaccgaa
     4861 caaggaatca caacggaatc ggagcttgcg aagaaagctc aaatcactcc gaacgacgtt
     4921 ttagcacttc cgggaatcac tcaaggtatc ttgctccctt tggatttcag tttaaaacat
     4981 aaatttagga ttcttatgct ccccatctgc gaacatctat aacatcgagt tcaccaagtt
     5041 ccaaatccgt gatctggaca ctgagcaagt gcttttcgag atcgccaaac cggagaacga
     5101 tcaggagaat gatgagtcgc cacaggagtc ggcaagatac gtgcgttata gatttgctcc
     5161 aaactttttg aaactcaaaa cggtcggagc aactgtggaa ttcaaagtag gagacatccc
     5221 aatccatcat ttccgaatga tcgaacgtca cttcttcaaa gatcgccttc tgaagtgttt
     5281 tgactttgaa ttcggattct gtattccgaa ttcacgaaac aactgtgaac atatctatga
     5341 gttccctcaa ctctctcaac aactcagtga gtcattattc taaaaagtac aattcaaaag
     5401 actaatctct ttttcagtgg acgacatgat caacaatcca aacgagactc gttctgacag
     5461 cttctatttc gtcgataaca aactcgtcat gcacaacaaa gccgactact catatgatgc
     5521 ataaatattt aatacaaaaa tgttctggat aattattctg tcgaatagaa aaaaaactcc
     5581 aaatgtgatt aaattccaat aattcctgtc tagtttgttc cttccccttc cccttctcat
     5641 gttcaatgca ttcctaagct tttcagttcc ccccttgttt ctatattttt tcgctgtcct
     5701 gtcacactcg ctaaaaacac taatcacacg gaaatctgtt ttcaataaaa actccaactt
     5761 taactcattt tcaatttcaa ctgaaagatt ttttcattag agaatgtcta gaactaggcc
     5821 cgggctacgt aatacgactc acttaaggcc ttgactagag ggtaccagag ctcacctagg
     5881 caggaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttgaagt
     5941 atactattta ttactcgata cttttgttca cataggtttt ttaaatcata ttttatgcat
     6001 catttatcat attcaatgca tcattcatat catagtcaat aaaaaggttg atttctcatg
     6061 ttctggtttc aaatgctgac tttggtaaaa agaacgcgtg cctgcctatt gcctatcttg
     6121 gcattttctc gataaatttt aaaatgtagg ttcgatctta tgagatttgt agtcaaaaga
     6181 gctcatatgt attcaggtag gtctggtagc gagaccaact taatagcatg acaagcattt
     6241 tcaatttgcc ctggagcgca attggttttt tattcgaaaa tcgcacattt ctgtttcccc
     6301 ataatataaa atttccagga cgatatatat tacattcttc acaaaatatt gcattacaga
     6361 caccgacaaa gaatctccac ctgatatgaa aacaatgagc caacaatgtt atctgtattg
     6421 ccaccaccca catttcctag tcattcagta tatattgttt caattgaatc attgcaggta
     6481 tatatcgaat tgaacttgta aggcttcatc ttcatttctc aatacatcat ccatcattcc
     6541 agagcagctc cggccacaca aaaattggtg gcggtctgat attgataatc gacttctttg
     6601 acgtgcctga cggagcagca aagcggagca ctgataagac aatgaagaac taaaaaattg
     6661 tcttcggttt tcagtcttta gttctgcagc actttatttt ttgtttctcc tatttttccg
     6721 cattttccta actttctgat gtccatttca aatgattttt gttataaaat tgtttaattt
     6781 cagggcgact aaaacctacc aaaacccata aaaaatgaca agtttgtaca aaaaagcagg
     6841 ctcaatggtg tcaaagggcg aagagctgtt tactggagtc gtcccaattc tggtggagct
     6901 ggatggagat gtcaacgggc ataagttttc agtgcgagga gagggagaag gagacgctac
     6961 caacggcaag ctgacactga aattcatcag caccacaggg aagctgcccg tgccttgggt
     7021 aagtttaaac atatatatac taactaaccc tgattattta aattttcagc caaccctggt
     7081 cactaccctg agttggggcg tccagtcatt tgcccgatac ccagaccaca tgaagcagca
     7141 tgatttcttt aaatccgcta tgcccgaggg ctacgtgcag gaacggacaa ttttctttaa
     7201 ggacgatgga acatacaaaa ctagtaagtt taaacagttc ggtactaact aaccatacat
     7261 atttaaattt tcaggagcag aggtgaagtt cgaaggagac actctggtca acaggatcga
     7321 gctgaagggc attgacttta aagaagatgg aaatatcctg ggccacaaac tggagtacaa
     7381 cgccatccat ggcaacgtgt acattacagc tgataagcag aaaaacggga ttaaggcaaa
     7441 tttcggactg aactgcaatg tggaagacgg gagtgtccag ctggccgatc actaccagca
     7501 gaacactcct atcggcgacg ggcccgtgct gctgcctgat aatcattatc tgagcaccca
     7561 gtccgtcctg tctaaggacc ccaatgagaa aagagatcac atggtgctgc tggagtttgt
     7621 gaccgctgct ggcattaccc tggggatgga cgagctgtac aagggaccag gcgagaattt
     7681 gtattttcag ggtcgtcttg tcaagttcct tgttggagga ccaggcatgg tgtctaaagg
     7741 cgaggaactg ttcactgggg tggtgcccat cctggtggag ctggacggcg atgtgaatgg
     7801 gcacaagttt agcgtgcgcg gagagggaga aggggacgct accaacggaa agctgacact
     7861 gaaactgatt tccaccacag gcaagctgcc cgtgccttgg ccaacactgg tgactaccct
     7921 gggatacggc ctccagagct tcgcccggta tccagaccac atgaagcagc atgatttctt
     7981 taaatcagct atgcccgagg gctacgtgca ggaacggaca atcttcttta aggacgatgg
     8041 gacatataaa actagagccg aggtgaagtt tgaaggggac actctggtga acaggatcga
     8101 gctgaaggga attgacttca aagaagatgg aaatatcctg ggccacaagc tggagtacaa
     8161 cttcaacagc cataacgtgt atatcaccgc cgataagcag aaaaacggca tcaaggctaa
     8221 tttcaaaatt aggcacaatg tggaagacgg cggggtgcag ctggctgatc attaccagca
     8281 gaacacacct atcggggacg gacccgtgct gctgcctgat aatcactacc tgagctatca
     8341 gtccgtgctg tctaaggacc caaacgagaa acgggatcat atggtgctga aagaatttgt
     8401 gactgccgct gggattaccc tgggaatgga tgagctgtat aagcacccag ctttcttgta
     8461 caaagtggga atgaagtggg cttacaagga ggagaacaac tttgagaagc gtcgtgccga
     8521 aggagacaag atccgcagaa agtacccaga ccgtattcca gtgattgttg agaaagcacc
     8581 aaagtcaaag ctccatgact tggataagaa gaagtacttg gtcccatccg atcttactgt
     8641 tggacagttc tacttcctca tcagaaaacg catccaactt cgtccagaag atgctctgtt
     8701 cttctttgtc aacaatgtca ttccacaaac catgaccaca atgggacaac tctaccaggt
     8761 aactaaccag tcgtttattt tcatttaatt aacccttttc tttattacag gaccatcacg
     8821 aggaagactt gttcctttac atcgcctaca gtgacgaaag tgtgtatgga ggagaggtcg
     8881 aaaagaagga ataaagtgtc atgtattatc cgccgacgaa tgtgtatact tgtacaatgg
     8941 aaatttcata cttctaatgg aaacccaaag cccctttaaa aaaatcttct cattttctgt
     9001 aataattctg tatttccttc acattttctt ctcatttttc tcgtatgacg gttccgagta
     9061 gtgacttcta ttcaaactat tgtctacgaa gttgctcttc tcccctccca gaaaatcatg
     9121 ttttctctcg gattttgata tttttttgct attactttca aatttttgat tcatgaataa
     9181 acgaagaaga cacaaaacca actttattat acatagttga taattcactg gccgtcgttt
     9241 tacactcgag acgtacggtg cgcgcgatgc attcgaagat ctgcccacta gtgagtcgta
     9301 ttataagtgc aagtaagatc agtgtttgtt tcgatatcag tctgtttcgt aacggtcttc
     9361 tgtataacta caaaaaagaa taaaaaaccg tatcacaact gagaaaaaag acaaaactta
     9421 ttgaacagtg ttcctttcaa aaaattattt tgtaaaaaat cgagtttttc tccaaaataa
     9481 cgcacgcaat caaaaacaag aaaactaacg tgttctcctg ggcgtggaac tccttatcaa
     9541 gattaccatt attgtctgtc agaattgata tttcaattat ttcatatcta gacaaacccg
     9601 ggtgtcttgc atttattagt actgtgagtt tacatacaaa cgaggcttac tcagataaca
     9661 tagagtcacc tctaattagc aaaaacccat ttctaattag ctatttcgac cagttctaaa
     9721 gtcaccgtaa ttagaaatgg tttttgtagc ttttctaatt agaaatgacc ccgagataag
     9781 tgaaatatgg cattttctgg gttttctagt aagaaatgat cccgcacgta gtgattccgc
     9841 gttacaagcc aaattcaaat taaaaaagta tattacacaa ttcaagttac attattttca
     9901 ggaaacaatg gcgaagagca aagcagagat tcaaaaggag gaaagagaca acagatatga
     9961 acgaagactt cttgaaaaga gaaagagaca tgaggtgagt aaccatgata ataaataatc
    10021 aagtaagggc tatagattta taaattctgg agttaacagt ttcgaaaaaa agtagttttc
    10081 atatgtgcga aggaatatca aaaatagggg aatagcctaa acgagtgaga aaattaaatg
    10141 tttgttattt tttcaaaaat gtctgcaggg ccgttatcct ttcaaatgca ctctatcaaa
    10201 catagatcat atccatcata tcttatccag aatttatatc atatccaaga ttgagagcgt
    10261 ttcaaggaaa atctcaaaca accatacaat tttgtagatc gacaacaagg cctccacttc
    10321 ttcatcgatg gcctctgcag cacctgcacc accaagtttg tatttttcat attaatataa
    10381 agtgagccac gcatagtcgt ttttaggatt tttgccagat cgatgtcgcg caaacagact
    10441 gttttttttt ctttgaaagc cgtgcgtgac atgggccacg gattgtaacg ccgcgctgga
    10501 tggataaaaa tttattcagt tttttagcct ttacaaaata tggttcgact actttcttta
    10561 tttttacatt tttataggtg ctcattgagc tgagatattt tgggtataat cggaaaatcg
    10621 ctgattttgg tcatttttct gcactaaaga ttacacaaaa agttcagtat ttagattcat
    10681 tgttgaaaat tggggatgat ctacagatac atatcgccgt gttcaaaaac a
//
