LOCUS       Exported                8453 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    PBMN YFP-ER50
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8453)
  AUTHORS   Miyazaki Y, Imoto H, Chen LC, Wandless TJ
  TITLE     Destabilizing domains derived from the human estrogen receptor.
  JOURNAL   J Am Chem Soc. 2012 Mar 7;134(9):3942-5. Epub 2012 Feb 22.
  PUBMED    22332638
REFERENCE   2  (bases 1 to 8453)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8453
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..1878
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1816..1832
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1896..1922
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1923..2639
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1968..1989)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2229..2248)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2576..2597
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3423..4008
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3590..3607)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3817..3836
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4009..4692
                     /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             4705..5391
                     /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             5440..6033
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6164..6181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6335..6923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6415..6434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7097..7957)
                     /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     7720..7739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     8025..8043
                     /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 acgtaccaag aacaactgga ccgaccggtg
     1681 gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc gccgacacca gactaagaac
     1741 ctagaacctc gctggaaagg accttacaca gtcctgctga ccacccccac cgccctcaaa
     1801 gtagacggca tcgcagcttg gatacacgcc gcccacgtga aggctgccga ccccgggggt
     1861 ggaccatcct ctagactgcc ggatccgcca ccatgtatcc gtacgacgta ccggattatg
     1921 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1981 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2041 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2101 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     2161 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2221 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2281 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2341 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2401 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2461 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2521 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2581 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     2641 aattcagcct tgccctgtca cttacagccg accagatggt ttccgcgctt ctcgacgctg
     2701 aacctccaat tctctattcc gaatacgacc caaccaggcc gttctccgag gcatctatga
     2761 tgggtctgct gacaaatctg gcagacaggg aactggtgca catgatcaat tgggcgaagc
     2821 gcgtacccgg attcgtcgat cttgcactcc atgatcaggt gcacttgctg gagtgcgctt
     2881 ggatggagat cctcatgatc gggctggtgt ggcggagtat ggaacacccc ggcaagttgc
     2941 tgtttgcgcc taacctcctg ttggacagga accaggggaa atgtgtggag ggcggtgtgg
     3001 aaatctttga catgctcctc gctacctcaa gccggtttag gatgatgaat ctgcagggcg
     3061 aagagttcgt gtgtctcaaa tctatcatac tgttgaacag cggagtctac accttcctct
     3121 ccagtactct gaaatctctg gaggagaaag atcatatcca tcgcgtgctg gacaagataa
     3181 ccgacacgtt gattcacttg atggccaaag ctgggctcac tctgcaacaa caacatcagc
     3241 gactggcaca gctgttgctg attttgagcc acattcggca catgtccagc aagagaatgg
     3301 agcacctcta tagtatgaag tgcaagaacg tcgtacccct gtcagatctg cttcttgaaa
     3361 tgcttgatgc ccaccggctg tgaatgcagg gccggcgcgc cgcggccgct acgtaaattc
     3421 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3481 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3541 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3601 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     3661 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     3721 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     3781 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     3841 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3901 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg
     3961 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcggc
     4021 ctgctgaagg agagtatgcg catcaagatg tacatggagg gcaccgtgaa cggccactac
     4081 ttcaagtgcg agggcgaggg cgacggcaac cccttcgccg gcacccagag catgagaatc
     4141 cacgtgaccg agggcgcccc cctgcccttc gccttcgaca tcctggcccc ctgctgcgag
     4201 tacggcagca ggaccttcgt gcaccacacc gccgagatcc ccgacttctt caagcagagc
     4261 ttccccgagg gcttcacctg ggagagaacc accacctacg aggacggcgg catcctgacc
     4321 gcccaccagg acaccagcct ggagggcaac tgcctgatct acaaggtgaa ggtgcacggc
     4381 accaacttcc ccgccgacgg ccccgtgatg aagaacaaga gcggcggctg ggagcccagc
     4441 accgaggtgg tgtaccccga gaacggcgtg ctgtgcggcc ggaacgtgat ggccctgaag
     4501 gtgggcgacc ggcacctgat ctgccaccac tacaccagct accggagcaa gaaggccgtg
     4561 cgcgccctga ccatgcccgg cttccacttc accgacatcc ggctccagat gctgcggaag
     4621 aagaaggacg agtacttcga gctgtacgag gccagcgtgg cccggtacag cgacctgccc
     4681 gagaaggcca acagatctcc cgggatggtg agcggcctgc tgaaggagag tatgcgcatc
     4741 aagatgtaca tggagggcac cgtgaacggc cactacttca agtgcgaggg cgagggcgac
     4801 ggcaacccct tcgccggcac ccagagcatg agaatccacg tgaccgaggg cgcccccctg
     4861 cccttcgcct tcgacatcct ggccccctgc tgcgagtacg gcagcaggac cttcgtgcac
     4921 cacaccgccg agatccccga cttcttcaag cagagcttcc ccgagggctt cacctgggag
     4981 agaaccacca cctacgagga cggcggcatc ctgaccgccc accaggacac cagcctggag
     5041 ggcaactgcc tgatctacaa ggtgaaggtg cacggcacca acttccccgc cgacggcccc
     5101 gtgatgaaga acaagagcgg cggctgggag cccagcaccg aggtggtgta ccccgagaac
     5161 ggcgtgctgt gcggccggaa cgtgatggcc ctgaaggtgg gcgaccggca cctgatctgc
     5221 caccactaca ccagctaccg gagcaagaag gccgtgcgcg ccctgaccat gcccggcttc
     5281 cacttcaccg acatccggct ccagatgctg cggaagaaga aggacgagta cttcgagctg
     5341 tacgaggcca gcgtggcccg gtacagcgac ctgcccgaga aggccaactg agtcgacgat
     5401 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     5461 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     5521 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     5581 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     5641 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     5701 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     5761 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     5821 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5881 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5941 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     6001 tgattgacta cccgtcagcg ggggtctttc attcacatgc agcatgtatc aaaattaatt
     6061 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     6121 cgcgcgggga gaggcggttt gcgtattggc gctcttccgc ttcctcgctc actgactcgc
     6181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7021 cctagatcct tttaaattaa aaatgaagtt tgcgcaaatc aatctaaagt atatatgagt
     7081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7681 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     7741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7981 gaagcattta tcagggttat tgtctcatga cattaaccta taaaaatagg cgtatcacga
     8041 ggccctttcg tcttcaagaa cagctttgct cttaggagtt tcctaataca tcccaaactc
     8101 aaatatataa agcatttgac ttgttctatg ccctaggggg cggggggaag ctaagccagc
     8161 tttttttaac atttaaaatg ttaattccat tttaaatgca cagatgtttt tatttcataa
     8221 gggtttcaat gtgcatgaat gctgcaatat tcctgttacc aaagctagta taaataaaaa
     8281 tagataaacg tggaaattac ttagagtttc tgtcattaac gtttccttcc tcagttgaca
     8341 acataaatgc gctgctgaga agccagtttg catctgtcag gatcaatttc ccattatgcc
     8401 agtcatatta attactagtc aattagttga tttttatttt tgacatatac atg
//
