LOCUS       sequence-164250-        8004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MMLV retroviral vector for expression of dGBP1-TagBFP with t-hcRed 
            as an infection/transfection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8004)
  AUTHORS   Tang JC, Drokhlyansky E, Etemad B, Rudolph S, Guo B, Wang S, Ellis 
            EG, Li JZ, Cepko CL
  TITLE     Detection and manipulation of live antigen-expressing cells using 
            conditionally stable nanobodies.
  JOURNAL   Elife. 2016 May 20;5. pii: e15312. doi: 10.7554/eLife.15312.
  PUBMED    27205882
REFERENCE   2  (bases 1 to 8004)
  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: "Elife."; 
            date: "2016-05-20"; volume: "5. pii: e15312. doi"; pages: " 
            10.7554/eLife.15312"
COMMENT     Sequence Label: pBMN-dGBP1-TagBFP
FEATURES             Location/Qualifiers
     source          1..8004
                     /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"
     regulatory      1887..1896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2250..2951
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    join(2973..3547,3548..3550,3551..3559)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3140..3157)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3367..3386
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3560..4243
                     /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             4256..4942
                     /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             4991..5584
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5715..5732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5886..6474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5966..5985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6648..7439,7440..7508))
                     /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     7271..7290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     7576..7594
                     /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 ccatggccga cgtgcagctc gtggaatccg
     1921 gaggagcact ggtccagcct gggggatcac tgagactgtc ttgtgtcgct tctggctttc
     1981 ctgtgaatcg gtactctatg agatggtaca ggcaggcacc tggaaaagag cgagaatggg
     2041 tcgccggaat gtcatctgct ggcgatagga gttcctacgt ggatagcgtg aaaggacggt
     2101 tcaccattag aagggacgat gctagaaaca ccgtgtacct ccagatgaac tccctgaaac
     2161 ctgaggacac tgctgtgtac tactacaacg tcaatgtggg ctttgaatac tggggacatg
     2221 gcacacaagt gactgttagt tttgcatgca tgagcgagct gattaaggag aacatgcaca
     2281 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     2341 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     2401 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     2461 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     2521 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     2581 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     2641 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     2701 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     2761 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     2821 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     2881 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     2941 agcttaatta agcggccgct acgtaaattc cgcccctctc cctccccccc ccctaacgtt
     3001 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     3061 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     3121 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     3181 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     3241 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     3301 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     3361 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     3421 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     3481 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     3541 tgataatatg gccacaacca tggtgagcgg cctgctgaag gagagtatgc gcatcaagat
     3601 gtacatggag ggcaccgtga acggccacta cttcaagtgc gagggcgagg gcgacggcaa
     3661 ccccttcgcc ggcacccaga gcatgagaat ccacgtgacc gagggcgccc ccctgccctt
     3721 cgccttcgac atcctggccc cctgctgcga gtacggcagc aggaccttcg tgcaccacac
     3781 cgccgagatc cccgacttct tcaagcagag cttccccgag ggcttcacct gggagagaac
     3841 caccacctac gaggacggcg gcatcctgac cgcccaccag gacaccagcc tggagggcaa
     3901 ctgcctgatc tacaaggtga aggtgcacgg caccaacttc cccgccgacg gccccgtgat
     3961 gaagaacaag agcggcggct gggagcccag caccgaggtg gtgtaccccg agaacggcgt
     4021 gctgtgcggc cggaacgtga tggccctgaa ggtgggcgac cggcacctga tctgccacca
     4081 ctacaccagc taccggagca agaaggccgt gcgcgccctg accatgcccg gcttccactt
     4141 caccgacatc cggctccaga tgctgcggaa gaagaaggac gagtacttcg agctgtacga
     4201 ggccagcgtg gcccggtaca gcgacctgcc cgagaaggcc aacagatctc ccgggatggt
     4261 gagcggcctg ctgaaggaga gtatgcgcat caagatgtac atggagggca ccgtgaacgg
     4321 ccactacttc aagtgcgagg gcgagggcga cggcaacccc ttcgccggca cccagagcat
     4381 gagaatccac gtgaccgagg gcgcccccct gcccttcgcc ttcgacatcc tggccccctg
     4441 ctgcgagtac ggcagcagga ccttcgtgca ccacaccgcc gagatccccg acttcttcaa
     4501 gcagagcttc cccgagggct tcacctggga gagaaccacc acctacgagg acggcggcat
     4561 cctgaccgcc caccaggaca ccagcctgga gggcaactgc ctgatctaca aggtgaaggt
     4621 gcacggcacc aacttccccg ccgacggccc cgtgatgaag aacaagagcg gcggctggga
     4681 gcccagcacc gaggtggtgt accccgagaa cggcgtgctg tgcggccgga acgtgatggc
     4741 cctgaaggtg ggcgaccggc acctgatctg ccaccactac accagctacc ggagcaagaa
     4801 ggccgtgcgc gccctgacca tgcccggctt ccacttcacc gacatccggc tccagatgct
     4861 gcggaagaag aaggacgagt acttcgagct gtacgaggcc agcgtggccc ggtacagcga
     4921 cctgcccgag aaggccaact gagtcgacga taaaataaaa gattttattt agtctccaga
     4981 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     5041 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     5101 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     5161 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     5221 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     5281 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     5341 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     5401 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     5461 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     5521 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     5581 cattcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa
     5641 atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5701 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5821 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5881 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5941 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6001 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6061 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6121 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6181 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6241 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6301 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6361 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6421 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6481 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6541 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6601 ttgcgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6661 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6721 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6781 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6841 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6901 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6961 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7021 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7081 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7141 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7201 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7261 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7321 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7381 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7441 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7501 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7561 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga acagctttgc
     7621 tcttaggagt ttcctaatac atcccaaact caaatatata aagcatttga cttgttctat
     7681 gccctagggg gcggggggaa gctaagccag ctttttttaa catttaaaat gttaattcca
     7741 ttttaaatgc acagatgttt ttatttcata agggtttcaa tgtgcatgaa tgctgcaata
     7801 ttcctgttac caaagctagt ataaataaaa atagataaac gtggaaatta cttagagttt
     7861 ctgtcattaa cgtttccttc ctcagttgac aacataaatg cgctgctgag aagccagttt
     7921 gcatctgtca ggatcaattt cccattatgc cagtcatatt aattactagt caattagttg
     7981 atttttattt ttgacatata catg
//
