LOCUS       Exported                5526 bp ds-DNA     circular SYN 23-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pmCherry-ATG5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5526)
  AUTHORS   Hamacher-Brady A, Brady NR, Logue SE, Sayen MR, Jinno M, Kirshenbaum
            LA, Gottlieb RA, Gustafsson AB
  TITLE     Response to myocardial ischemia/reperfusion injury involves Bnip3 
            and autophagy.
  JOURNAL   Cell Death Differ. 2006 Apr 28. ():.
  PUBMED    16645637
REFERENCE   2  (bases 1 to 5526)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5526
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(2..21)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(293..397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      424..879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(511..530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     721..742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    886..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(897..916)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     951..970
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2007..2714)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2282..2301)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2551..2569
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2753..2956)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2782..2802)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2957..3260
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3435..4023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3515..3534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4352..4399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4424..4443
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4631..5425)
                     /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(4742..4761)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5352..5371
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5460..291)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5474..83
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
      301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgcgttaaga tacattgatg
      901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1081 acctctacaa atgtggtatg gctgattatg atcagttatc tagatccggt ggatcctcaa
     1141 tctgttggct gggggacaat gctaatatga agaaagttat ctgggtagct cagatgctcg
     1201 ctcagccact gcagaggggt ttccagcatt ggctctatcc cgtgaatcat cacctggctc
     1261 ctcttctctc catcttcagg ggcgactgcg gaaggacaga cttctctgag gagatctcca
     1321 agtgtgtgca gctgtccatc tgcggccaca ggccggaaca gcttctggat gaaaggccgc
     1381 tccgtcgtgg tctgatatat tctaaagggg atataacgaa atccattttc ttctggagga
     1441 tattccatga gtttccggtt gatggcccaa aactggtcaa atctgtcatt ctgcagtccc
     1501 atccagagct gcttgtggtc ttttttctgc atttcgttga tcacttgact tttatgcttt
     1561 aaagcatcag cttctttcat acacgacata aagtgagcct caaccgcatc cttggatgga
     1621 cagtgtagaa ggtccttttc tggaaaactc ttgaaatgta ctgtgatgtt ccaaggaaga
     1681 gctgaacttg atgcaagaag atcaaatagt aaaccaattg gataatgcca tttcaggggt
     1741 gtgccttcat attcaaacca tatctcacta acatcttctt gtctcataac cttctgaaag
     1801 tgctttttca ctttgtcagt taccaacgtc aaatagctga ctcttggcaa aagcaaatag
     1861 tatggttctg cttctctttc agttatctca tcctgataga gagtaaagca agttggaatt
     1921 cgtccaaacc acacatctcg aagcacatct ttgtcatctg tcatggtacc gtcgactgca
     1981 gaattcgaag cttgagctcg agatctcttg tacagctcgt ccatgccgcc ggtggagtgg
     2041 cggccctcgg cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg
     2101 tccaacttga tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag
     2161 gtggtcttga cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc
     2221 tcgcccttca gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg
     2281 gtcttcttct gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg
     2341 tagatgaact cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc
     2401 tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg
     2461 gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg
     2521 atgtcccagg cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg
     2581 ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcacggag
     2641 ccctccatgt gcaccttgaa gcgcatgaac tccttgatga tggccatgtt atcctcctcg
     2701 cccttgctca ccattaccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc
     2761 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     2821 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     2881 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     2941 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     3001 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     3061 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     3121 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     3181 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     3241 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     3301 ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     3361 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3601 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3841 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     4201 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     4261 gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     4321 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     4381 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     4441 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     4501 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     4561 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     4621 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4681 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4741 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4801 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     4861 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     4921 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4981 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     5041 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     5101 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     5161 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     5221 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     5281 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     5341 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     5401 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     5461 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     5521 gaggcg
//
