LOCUS       pcDNA3-Lyso-mChe        7962 bp DNA     circular SYN 20-SEP-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7962)
  AUTHORS   Jin Zhang Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2022 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     regulatory      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             907..2172
                     /codon_start=1
                     /label=LAMP1
                     /note="note=""direct strand 
                     translation""translation=""KLAAATMAAPGSARRPLLLLLLLLLLGLMHCA
                     SAAMFMVKNGNGTACIMANFSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENT
                     SDPSLVIAFGRGHTLTLNFTRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITD
                     IRADIDKKYRCVSGTQVHMNNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPP
                     APPSPSPSPVPKSPSVDKYNVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNK
                     TSASGSCGAHLVTLELHSEGTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAA
                     NGSLRALQATVGNSYKCNAEEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDEN
                     SMLIPIAVGGALAGLVLIVLIAYLVGRKRSHAGYQTIGGGGG"""
                     /note="color: #800040"
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTIGGGGG"
     CDS             2182..2889
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMXELYK"
     CDS             2899..3453
                     /codon_start=1
                     /label=Rheb (C181S)
                     /note="color: #993366"
                     /translation="MPQSKSRKIAILGYRSVGKSSLTIQFVEGQFVDSYDPTIENTFTK
                     LITVNGQEYHLQLVDTAGQDEYSIFPQTYSIDINGYILVYSVTSIKSFEVIKVIHGKLL
                     DMVGKVQIPIMLVGNKKDLHMERVISYEEGKALAESWNAAFLESSAKENQTAVDVFRRI
                     ILEAEKMDGAASQGKSSSSVM"
     promoter        complement(3514..3532)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    3558..3782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3828..4256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4270..4600
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4451..4586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4667..5461
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5635..5756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5805..5821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5829..5845
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(5853..5883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5853 .. 5859 / #ffffff / -10
                        2: 5860 .. 5877 / #ffffff
                        3: 5878 .. 5883 / #ffffff / -35"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(6207..6795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6966..7826)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6966 .. 7757 / #ccffcc
                        2: 7758 .. 7826 / #ccffcc / signal sequence
                     Cleavage site after base 7757"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7827..7931)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gccaccatgg cggcccccgg cagcgcccgg cgacccctgc tgctgctact gctgttgctg
      961 ctgctcggcc tcatgcattg tgcgtcagca gcaatgttta tggtgaaaaa tggcaacggg
     1021 accgcgtgca taatggccaa cttctctgct gccttctcag tgaactacga caccaagagt
     1081 ggccctaaga acatgacctt tgacctgcca tcagatgcca cagtggtgct caaccgcagc
     1141 tcctgtggaa aagagaacac ttctgacccc agtctcgtga ttgcttttgg aagaggacat
     1201 acactcactc tcaatttcac gagaaatgca acacgttaca gcgtccagct catgagtttt
     1261 gtttataact tgtcagacac acaccttttc cccaatgcga gctccaaaga aatcaagact
     1321 gtggaatcta taactgacat cagggcagat atagataaaa aatacagatg tgttagtggc
     1381 acccaggtcc acatgaacaa cgtgaccgta acgctccatg atgccaccat ccaggcgtac
     1441 ctttccaaca gcagcttcag caggggagag acacgctgtg aacaagacag gccttcccca
     1501 accacagcgc cccctgcgcc acccagcccc tcgccctcac ccgtgcccaa gagcccctct
     1561 gtggacaagt acaacgtgag cggcaccaac gggacctgcc tgctggccag catggggctg
     1621 cagctgaacc tcacctatga gaggaaggac aacacgacgg tgacaaggct tctcaacatc
     1681 aaccccaaca agacctcggc cagcgggagc tgcggcgccc acctggtgac tctggagctg
     1741 cacagcgagg gcaccaccgt cctgctcttc cagttcggga tgaatgcaag ttctagccgg
     1801 tttttcctac aaggaatcca gttgaataca attcttcctg acgccagaga ccctgccttt
     1861 aaagctgcca acggctccct gcgagcgctg caggccacag tcggcaattc ctacaagtgc
     1921 aacgcggagg agcacgtccg tgtcacgaag gcgttttcag tcaatatatt caaagtgtgg
     1981 gtccaggctt tcaaggtgga aggtggccag tttggctctg tggaggagtg tctgctggac
     2041 gagaacagca tgctgatccc catcgctgtg ggtggtgccc tggcggggct ggtcctcatc
     2101 gtcctcatcg cctacctcgt cggcaggaag aggagtcacg caggctacca gactatcggt
     2161 ggtggcggtg gtgcggatcc catggtgagc aagggcgagg aggataacat ggccatcatc
     2221 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2281 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2341 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2401 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     2461 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     2521 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     2581 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     2641 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2701 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2761 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2821 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggncgag
     2881 ctgtacaagc gcatgcacat gccgcagtcc aagtcccgga agatcgcgat cctgggctac
     2941 cggtctgtgg ggaaatcctc attgacgatt caatttgttg aaggccaatt tgtggactcc
     3001 tacgatccaa ccatagaaaa cacttttaca aagttgatca cagtaaatgg acaagaatat
     3061 catcttcaac ttgtagacac agccgggcaa gatgaatatt ctatctttcc tcagacatac
     3121 tccatagata ttaatggcta tattcttgtg tattctgtta catcaatcaa aagttttgaa
     3181 gtgattaaag ttatccatgg caaattgttg gatatggtgg ggaaagtaca aatacctatt
     3241 atgttggttg ggaataagaa agacctgcat atggaaaggg tgatcagtta tgaagaaggg
     3301 aaagctttgg cagaatcttg gaatgcagct tttttggaat cttctgctaa agaaaatcag
     3361 actgctgtgg atgtttttcg aaggataatt ttggaggcag aaaaaatgga cggggcagct
     3421 tcacaaggca agtcttcatc ctcggtgatg tgagaattct gcagatatcc atcacactgg
     3481 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3541 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3661 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3721 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3781 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     4201 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4261 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4381 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4441 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4501 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4561 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4621 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4681 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4741 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4801 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4861 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4921 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4981 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5041 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5101 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5161 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5221 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5281 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5341 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5401 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5461 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5521 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5581 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5641 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5701 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5761 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
