LOCUS       sequence-46848-e        8462 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 8462)
  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 8462)
  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: "J Am Chem 
            Soc."; date: "2012-03-7"; volume: "134(9)"; pages: "3942-5. Epub 
            2012 Feb 22"
COMMENT     Sequence Label: pBMN ER50-YFP
FEATURES             Location/Qualifiers
     source          1..8462
                     /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..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             join(2637..2639,2640..3350)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2679..2700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2940..2959)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3287..3308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3357..3383
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(3432..4005,4006..4008,4009..4017)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3599..3616)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3826..3845
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4018..4701
                     /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             4714..5400
                     /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             5449..6042
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6173..6190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6344..6932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6424..6443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7106..7897,7898..7966))
                     /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     7729..7748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     8034..8052
                     /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 ccatgagcct tgccctgtca cttacagccg
     1921 accagatggt ttccgcgctt ctcgacgctg aacctccaat tctctattcc gaatacgacc
     1981 caaccaggcc gttctccgag gcatctatga tgggtctgct gacaaatctg gcagacaggg
     2041 aactggtgca catgatcaat tgggcgaagc gcgtacccgg attcgtcgat cttgcactcc
     2101 atgatcaggt gcacttgctg gagtgcgctt ggatggagat cctcatgatc gggctggtgt
     2161 ggcggagtat ggaacacccc ggcaagttgc tgtttgcgcc taacctcctg ttggacagga
     2221 accaggggaa atgtgtggag ggcggtgtgg aaatctttga catgctcctc gctacctcaa
     2281 gccggtttag gatgatgaat ctgcagggcg aagagttcgt gtgtctcaaa tctatcatac
     2341 tgttgaacag cggagtctac accttcctct ccagtactct gaaatctctg gaggagaaag
     2401 atcatatcca tcgcgtgctg gacaagataa ccgacacgtt gattcacttg atggccaaag
     2461 ctgggctcac tctgcaacaa caacatcagc gactggcaca gctgttgctg attttgagcc
     2521 acattcggca catgtccagc aagagaatgg agcacctcta tagtatgaag tgcaagaacg
     2581 tcgtacccct gtcagatctg cttcttgaaa tgcttgatgc ccaccggctg gaattcgtga
     2641 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2701 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2761 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2821 ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     2881 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2941 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3001 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3061 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3121 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3181 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3241 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3301 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag gtcgactatc
     3361 cgtacgacgt accagactac gcactcgact aactcgaggg ccggcgcgcc gcggccgcta
     3421 cgtaaattcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3481 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3541 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3601 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3661 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3721 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3781 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3841 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3901 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc
     3961 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     4021 gtgagcggcc tgctgaagga gagtatgcgc atcaagatgt acatggaggg caccgtgaac
     4081 ggccactact tcaagtgcga gggcgagggc gacggcaacc ccttcgccgg cacccagagc
     4141 atgagaatcc acgtgaccga gggcgccccc ctgcccttcg ccttcgacat cctggccccc
     4201 tgctgcgagt acggcagcag gaccttcgtg caccacaccg ccgagatccc cgacttcttc
     4261 aagcagagct tccccgaggg cttcacctgg gagagaacca ccacctacga ggacggcggc
     4321 atcctgaccg cccaccagga caccagcctg gagggcaact gcctgatcta caaggtgaag
     4381 gtgcacggca ccaacttccc cgccgacggc cccgtgatga agaacaagag cggcggctgg
     4441 gagcccagca ccgaggtggt gtaccccgag aacggcgtgc tgtgcggccg gaacgtgatg
     4501 gccctgaagg tgggcgaccg gcacctgatc tgccaccact acaccagcta ccggagcaag
     4561 aaggccgtgc gcgccctgac catgcccggc ttccacttca ccgacatccg gctccagatg
     4621 ctgcggaaga agaaggacga gtacttcgag ctgtacgagg ccagcgtggc ccggtacagc
     4681 gacctgcccg agaaggccaa cagatctccc gggatggtga gcggcctgct gaaggagagt
     4741 atgcgcatca agatgtacat ggagggcacc gtgaacggcc actacttcaa gtgcgagggc
     4801 gagggcgacg gcaacccctt cgccggcacc cagagcatga gaatccacgt gaccgagggc
     4861 gcccccctgc ccttcgcctt cgacatcctg gccccctgct gcgagtacgg cagcaggacc
     4921 ttcgtgcacc acaccgccga gatccccgac ttcttcaagc agagcttccc cgagggcttc
     4981 acctgggaga gaaccaccac ctacgaggac ggcggcatcc tgaccgccca ccaggacacc
     5041 agcctggagg gcaactgcct gatctacaag gtgaaggtgc acggcaccaa cttccccgcc
     5101 gacggccccg tgatgaagaa caagagcggc ggctgggagc ccagcaccga ggtggtgtac
     5161 cccgagaacg gcgtgctgtg cggccggaac gtgatggccc tgaaggtggg cgaccggcac
     5221 ctgatctgcc accactacac cagctaccgg agcaagaagg ccgtgcgcgc cctgaccatg
     5281 cccggcttcc acttcaccga catccggctc cagatgctgc ggaagaagaa ggacgagtac
     5341 ttcgagctgt acgaggccag cgtggcccgg tacagcgacc tgcccgagaa ggccaactga
     5401 gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     5461 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     5521 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     5581 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     5641 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     5701 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     5761 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     5821 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     5881 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     5941 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     6001 tcctctgagt gattgactac ccgtcagcgg gggtctttca ttcacatgca gcatgtatca
     6061 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     6121 atcggccaac gcgcggggag aggcggtttg cgtattggcg ctcttccgct tcctcgctca
     6181 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6241 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6781 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6961 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7021 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt gcgcaaatca atctaaagta
     7081 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7141 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7201 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7261 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7321 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7381 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7441 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7501 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7561 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7621 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7681 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     7741 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7801 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7861 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7921 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7981 aatattattg aagcatttat cagggttatt gtctcatgac attaacctat aaaaataggc
     8041 gtatcacgag gccctttcgt cttcaagaac agctttgctc ttaggagttt cctaatacat
     8101 cccaaactca aatatataaa gcatttgact tgttctatgc cctagggggc ggggggaagc
     8161 taagccagct ttttttaaca tttaaaatgt taattccatt ttaaatgcac agatgttttt
     8221 atttcataag ggtttcaatg tgcatgaatg ctgcaatatt cctgttacca aagctagtat
     8281 aaataaaaat agataaacgt ggaaattact tagagtttct gtcattaacg tttccttcct
     8341 cagttgacaa cataaatgcg ctgctgagaa gccagtttgc atctgtcagg atcaatttcc
     8401 cattatgcca gtcatattaa ttactagtca attagttgat ttttattttt gacatataca
     8461 tg
//
