LOCUS       sequence-358076-        6414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral transfer vector expressing TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  TITLE     Retroviral TagBFP expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6414)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMIBlu
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             323..839
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    903..1244
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1311..1727
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1657..1679
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1695..1711
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1756..2323
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1773..2325
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1917..1934)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2144..2163
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2340..3041
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     LTR             3131..3645
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3807..3823
                     /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(join(3831..3837,3838..3855,3856..3861))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3876..3897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4014..4031)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4185..4773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4265..4284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4944..5735,5736..5804))
                     /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     5567..5586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5805..5909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5977..5995
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6033..6055)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6155..6174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6368..6390
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
       61 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      121 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      181 ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc
      241 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
      301 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
      361 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
      421 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
      481 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
      541 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      601 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      661 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
      721 gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg
      781 ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt
      841 ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag
      901 gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca
      961 tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc
     1021 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
     1081 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
     1141 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     1201 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
     1261 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
     1321 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1381 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1441 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1501 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1561 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1621 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1681 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga attagatctc
     1741 tcgaggttaa cgaattccgc cccccccccc taacgttact ggccgaagcc gcttggaata
     1801 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     1861 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     1921 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     1981 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     2041 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     2101 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     2161 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     2221 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     2281 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatga
     2341 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     2401 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2461 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     2521 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     2581 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2641 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     2701 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     2761 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     2821 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
     2881 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     2941 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     3001 actgcgacct ccctagcaaa ctggggcaca agcttaatta agaattatca agcttatcga
     3061 taccgtcgac ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa
     3121 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     3181 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     3241 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3301 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     3361 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3421 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     3481 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     3541 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     3601 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     3661 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     3721 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     3781 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     3841 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3901 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3961 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4021 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4081 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5941 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6001 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6061 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6121 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6181 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6241 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6301 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6361 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgca
//
