LOCUS       Exported                9306 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMSCV-OPA1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9306)
  TITLE     David CHan plasmids
REFERENCE   2  (bases 1 to 9306)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9306
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     146..165
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             234..833
                     /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(234..253)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     730..750
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             933..1447
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    1609..1625
                     /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)."
     promoter        complement(1633..1663)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1678..1699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1816..1833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1987..2575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2067..2086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2746..3606)
                     /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     3369..3388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3607..3711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3779..3797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3835..3857)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3957..3976
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4170..4192
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4320..4339
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             4539..5055
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5119..5460
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             5527..5943
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5873..5895
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5911..5927
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     polyA_signal    8885..9006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8896..8915)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8950..8969
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        9020..213
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9055..9078)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
       61 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
      121 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
      181 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg
      241 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
      301 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
      361 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
      421 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
      481 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
      541 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
      601 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
      661 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
      721 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
      781 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
      841 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg ataaaataaa
      901 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
      961 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     1021 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     1081 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     1141 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     1201 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     1261 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     1321 gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     1381 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     1441 ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat
     1501 attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga
     1561 aatagttcat tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct
     1621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1741 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3721 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3781 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3841 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3901 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3961 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4021 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4081 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4141 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca
     4201 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     4261 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     4321 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     4381 atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag
     4441 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
     4501 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     4561 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     4621 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     4681 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     4741 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4801 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     4861 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     4921 tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat
     4981 ccgaatcgtg gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct
     5041 cgggggtctt tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg
     5101 acccccccgc cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg
     5161 cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc
     5221 tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg
     5281 agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg
     5341 tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc
     5401 ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg
     5461 cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa
     5521 aattagggcc agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc
     5581 gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc
     5641 tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac
     5701 ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag
     5761 gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc
     5821 tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa
     5881 cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc
     5941 gccggaatta gatctctcga gaattcacgc gtggtaccat gtggcgacta cgtcgggccg
     6001 ctgtggcctg tgaggtctgc cagtctttag tgaaacacag ctctggaata aaaggaagtt
     6061 taccactaca aaaactacat ctggtttcac gaagcattta tcattcacat catcctacct
     6121 taaagcttca acgaccccaa ttaaggacat cctttcagca gttctcttct ctgacaaacc
     6181 ttcctttacg taaactgaaa ttctctccaa ttaaatatgg ctaccagcct cgcaggaatt
     6241 tttggccagc aagattagct acgagactct taaaacttcg ctatctcata ctaggatcgg
     6301 ctgttggggg tggctacaca gccaaaaaga cttttgatca gtggaaagat atgataccgg
     6361 accttagtga atataaatgg attgtgcctg acattgtgtg ggaaattgat gagtatatcg
     6421 attttgagaa aattagaaaa gcccttccta attcagaaga ccttgtaaag ttagcaccag
     6481 actttgacaa gattgttgaa agccttagct tattgaagga cttttttacc tcaggttctc
     6541 cggaagaaac ggcgtttaga gcaacagatc gtggatctga aagtgacaag cattttagaa
     6601 aggtgtcaga caaagagaaa attgaccaac ttcaggaaga acttctgcac actcagttga
     6661 agtatcagag aatcttggaa cgattagaaa aggagaacaa agaattgaga aaattagtat
     6721 tgcagaaaga tgacaaaggc attcatcata gaaagcttaa gaaatctttg attgacatgt
     6781 attctgaagt tcttgatgtt ctctctgatt atgatgccag ttataatacg caagatcatc
     6841 tgccacgggt tgttgtggtt ggagatcaga gtgctggaaa gactagtgtg ttggaaatga
     6901 ttgcccaagc tcgaatattc ccaagaggat ctggggagat gatgacacgt tctccagtta
     6961 aggtgactct gagtgaaggt cctcaccatg tggccctatt taaagatagt tctcgggagt
     7021 ttgatcttac caaagaagaa gatcttgcag cattaagaca tgaaatagaa cttcgaatga
     7081 ggaaaaatgt gaaagaaggc tgtaccgtta gccctgagac catatcctta aatgtaaaag
     7141 gccctggact acagaggatg gtgcttgttg acttaccagg tgtgattaat actgtgacat
     7201 caggcatggc tcctgacaca aaggaaacta ttttcagtat cagcaaagct tacatgcaga
     7261 atcctaatgc catcatactg tgtattcaag atggatctgt ggatgctgaa cgcagtattg
     7321 ttacagactt ggtcagtcaa atggaccctc atggaaggag aaccatattc gttttgacca
     7381 aagtagacct ggcagagaaa aatgtagcca gtccaagcag gattcagcag ataattgaag
     7441 gaaagctctt cccaatgaaa gctttaggtt attttgctgt tgtaacagga aaagggaaca
     7501 gctctgaaag cattgaagct ataagagaat atgaagaaga gttttttcag aattcaaagc
     7561 tcctaaagac aagcatgcta aaggcacacc aagtgactac aagaaattta agccttgcag
     7621 tatcagactg cttttggaaa atggtacgag agtctgttga acaacaggct gatagtttca
     7681 aagcaacacg ttttaacctt gaaactgaat ggaagaataa ctatcctcgc ctgcgggaac
     7741 ttgaccggaa tgaactattt gaaaaagcta aaaatgaaat ccttgatgaa gttatcagtc
     7801 tgagccaggt tacaccaaaa cattgggagg aaatccttca acaatctttg tgggaaagag
     7861 tatcaactca tgtgattgaa aacatctacc ttccagctgc gcagaccatg aattcaggaa
     7921 cttttaacac cacagtggat atcaagctta aacagtggac tgataaacaa cttcctaata
     7981 aagcagtaga ggttgcttgg gagaccctac aagaagaatt ttcccgcttt atgacagaac
     8041 cgaaagggaa agagcatgat gacatatttg ataaacttaa agaggccgtt aaggaagaaa
     8101 gtattaaacg acacaagtgg aatgactttg cggaggacag cttgagggtt attcaacaca
     8161 atgctttgga agaccgatcc atatctgata aacagcaatg ggatgcagct atttatttta
     8221 tggaagaggc tctgcaggct cgtctcaagg atactgaaaa tgcaattgaa aacatggtgg
     8281 gtccagactg gaaaaagagg tggttatact ggaagaatcg gacccaagaa cagtgtgttc
     8341 acaatgaaac caagaatgaa ttggagaaga tgttgaaatg taatgaggag cacccagctt
     8401 atcttgcaag tgatgaaata accacagtcc ggaagaacct tgaatcccga ggagtagaag
     8461 tagatccaag cttgattaag gatacttggc atcaagttta tagaagacat tttttaaaaa
     8521 cagctctaaa ccattgtaac ctttgtcgaa gaggttttta ttactaccaa aggcattttg
     8581 tagattctga gttggaatgc aatgatgtgg tcttgttttg gcgtatacag cgcatgcttg
     8641 ctatcaccgc aaatacttta aggcaacaac ttacaaatac tgaagttagg cgattagaga
     8701 aaaatgttaa agaggtattg gaagattttg ctgaagatgg tgagaagaag attaaattgc
     8761 ttactggtaa acgcgttcaa ctggcggaag acctcaagaa agttagagaa attcaagaaa
     8821 aacttgatgc tttcattgaa gctcttcatc aggagaaata agcggccgct tcgagcagac
     8881 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     8941 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     9001 caagttaacg aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     9061 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     9121 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     9181 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     9241 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     9301 tggaag
//
