LOCUS       sequence-112588-        8181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  AUTHORS   Iwamoto M, Bjoerklund T, Lundberg C, Kirik D, Wandless TJ
  TITLE     A general chemical method to regulate protein stability in the 
            mammalian central nervous system.
  JOURNAL   Chem Biol. 2010 Sep 24. 17(9):981-8.
  PUBMED    20851347
REFERENCE   2  (bases 1 to 8181)
  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: "Chem Biol. 
            2010 Sep 24. 17(9):981-8."
COMMENT     Sequence Label: pBMN DHFR(DD)-YFP
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1079..1495
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1425..1447
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1463..1479
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1505..1879
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1817..1833
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1894..2370
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             join(2377..2379,2380..3090)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2419..2440)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2680..2699)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3027..3048
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3097..3123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(3150..3724,3725..3727,3728..3736)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3317..3334)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3544..3563
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3737..4420
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     CDS             4433..5119
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     LTR             5168..5761
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5892..5909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6063..6651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6143..6162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6825..7616,7617..7685))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7448..7467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     7753..7771
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaagc ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt
      901 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg
     1081 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1141 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1201 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1261 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1321 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1381 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1441 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1501 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1561 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1621 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1681 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     1741 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     1801 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     1861 tggaccatcc tctagactgc cggatccgcc accatgatca gtctgattgc ggcgttagcg
     1921 gtagattacg ttatcggcat ggaaaacgcc atgccgtgga acctgcctgc cgatctcgcc
     1981 tggtttaaac gcaacacctt aaataaaccc gtgattatgg gccgccatac ctgggaatca
     2041 atcggtcgtc cgttgccagg acgcaaaaat attatcctca gcagtcaacc gagtacggac
     2101 gatcgcgtaa cgtgggtgaa gtcggtggat gaagccatcg cggcgtgtgg tgacgtacca
     2161 gaaatcatgg tgattggcgg cggtcgcgtt attgaacagt tcttgccaaa agcgcaaaaa
     2221 ctgtatctga cgcatatcga cgcagaagtg gaaggcgaca cccatttccc ggattacgag
     2281 ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca gaactctcac
     2341 agctattgct ttgagattct ggagcggcga gcatgcgtga gcaagggcga ggagctgttc
     2401 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2461 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2521 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2581 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2641 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2701 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2761 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2821 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2881 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2941 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     3001 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3061 atcactctcg gcatggacga gctgtacaag gtcgactatc cgtacgacgt accagactac
     3121 gcatgaccgc ggccgctacg taaattccgc ccctctccct cccccccccc taacgttact
     3181 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     3241 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3301 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3361 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3421 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3481 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3541 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3601 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3661 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3721 taatatggcc acaaccatgg tgagcggcct gctgaaggag agtatgcgca tcaagatgta
     3781 catggagggc accgtgaacg gccactactt caagtgcgag ggcgagggcg acggcaaccc
     3841 cttcgccggc acccagagca tgagaatcca cgtgaccgag ggcgcccccc tgcccttcgc
     3901 cttcgacatc ctggccccct gctgcgagta cggcagcagg accttcgtgc accacaccgc
     3961 cgagatcccc gacttcttca agcagagctt ccccgagggc ttcacctggg agagaaccac
     4021 cacctacgag gacggcggca tcctgaccgc ccaccaggac accagcctgg agggcaactg
     4081 cctgatctac aaggtgaagg tgcacggcac caacttcccc gccgacggcc ccgtgatgaa
     4141 gaacaagagc ggcggctggg agcccagcac cgaggtggtg taccccgaga acggcgtgct
     4201 gtgcggccgg aacgtgatgg ccctgaaggt gggcgaccgg cacctgatct gccaccacta
     4261 caccagctac cggagcaaga aggccgtgcg cgccctgacc atgcccggct tccacttcac
     4321 cgacatccgg ctccagatgc tgcggaagaa gaaggacgag tacttcgagc tgtacgaggc
     4381 cagcgtggcc cggtacagcg acctgcccga gaaggccaac agatctcccg ggatggtgag
     4441 cggcctgctg aaggagagta tgcgcatcaa gatgtacatg gagggcaccg tgaacggcca
     4501 ctacttcaag tgcgagggcg agggcgacgg caaccccttc gccggcaccc agagcatgag
     4561 aatccacgtg accgagggcg cccccctgcc cttcgccttc gacatcctgg ccccctgctg
     4621 cgagtacggc agcaggacct tcgtgcacca caccgccgag atccccgact tcttcaagca
     4681 gagcttcccc gagggcttca cctgggagag aaccaccacc tacgaggacg gcggcatcct
     4741 gaccgcccac caggacacca gcctggaggg caactgcctg atctacaagg tgaaggtgca
     4801 cggcaccaac ttccccgccg acggccccgt gatgaagaac aagagcggcg gctgggagcc
     4861 cagcaccgag gtggtgtacc ccgagaacgg cgtgctgtgc ggccggaacg tgatggccct
     4921 gaaggtgggc gaccggcacc tgatctgcca ccactacacc agctaccgga gcaagaaggc
     4981 cgtgcgcgcc ctgaccatgc ccggcttcca cttcaccgac atccggctcc agatgctgcg
     5041 gaagaagaag gacgagtact tcgagctgta cgaggccagc gtggcccggt acagcgacct
     5101 gcccgagaag gccaactgag tcgacgataa aataaaagat tttatttagt ctccagaaaa
     5161 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     5221 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     5281 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     5341 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     5401 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     5461 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     5521 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     5581 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     5641 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     5701 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     5761 tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg
     5821 gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattggcgc
     5881 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5941 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6001 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6061 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6481 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6661 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6721 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttg
     6781 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7441 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgaca
     7741 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaaca gctttgctct
     7801 taggagtttc ctaatacatc ccaaactcaa atatataaag catttgactt gttctatgcc
     7861 ctagggggcg gggggaagct aagccagctt tttttaacat ttaaaatgtt aattccattt
     7921 taaatgcaca gatgttttta tttcataagg gtttcaatgt gcatgaatgc tgcaatattc
     7981 ctgttaccaa agctagtata aataaaaata gataaacgtg gaaattactt agagtttctg
     8041 tcattaacgt ttccttcctc agttgacaac ataaatgcgc tgctgagaag ccagtttgca
     8101 tctgtcagga tcaatttccc attatgccag tcatattaat tactagtcaa ttagttgatt
     8161 tttatttttg acatatacat g
//
