LOCUS       sequence-203918-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  creating stable line with RFP and BSD resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  AUTHORS   Liao J, Wei Q, Fan J, Zou Y, Song D, Liu J, Liu F, Ma C, Hu X, Li L,
            Yu Y, Qu X, Chen L, Yu X, Zhang Z, Zhao C, Zeng Z, Zhang R, Yan S, 
            Wu T, Wu X, Shu Y, Lei J, Li Y, Zhang W, Wang J, Reid RR, Lee MJ, 
            Huang W, Wolf JM, He TC, Wang J
  TITLE     Characterization of retroviral infectivity and superinfection 
            resistance during retrovirus-mediated transduction of mammalian 
            cells.
  JOURNAL   Gene Ther. 2017 Jun;24(6):333-341. doi: 10.1038/gt.2017.24. Epub 
            2017 May 4.
  PUBMED    28387759
REFERENCE   2  (bases 1 to 6789)
  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"; doi: 
            "10.1038/gt.2017.24"; journalName: "Gene Ther"; date: "2017-06"; 
            volume: "24"; issue: "6"; pages: "333-341"
COMMENT     Sequence Label: pRV-RB
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    73..414
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             481..897
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     827..849
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     865..881
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             908..1306
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        1340..1551
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1568..1836
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1879..2556
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1879..2556
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2009..2027)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2277..2296
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2471..2494
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2590..2613
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             2676..3190
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3352..3368
                     /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(3376..3382,3383..3400,3401..3406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3421..3442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3559..3576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3730..4318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3810..3829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4489..5280,5281..5349))
                     /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     5112..5131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5350..5454)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5522..5540
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5578..5600)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5700..5719
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5913..5935
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6063..6082
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6282..6789,1..9)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
       61 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
      121 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
      181 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
      241 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
      301 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
      361 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
      421 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
      481 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
      541 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      601 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      661 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      721 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      781 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      841 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      901 attcaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
      961 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     1021 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     1081 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     1141 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     1201 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     1261 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagttt aaacatgcag
     1321 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     1381 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     1441 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     1501 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagcta
     1561 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     1621 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     1681 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     1741 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     1801 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacatc
     1861 gatccgcgga gatccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc
     1921 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1981 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     2041 cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag
     2101 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2161 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2221 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2281 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac
     2341 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac
     2401 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag
     2461 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2521 gagcgcgccg agggccgcca ctccaccggc gcctaagtcg acggatccac gcgtgggccc
     2581 ggaggcggcg attacaagga tgacgacgat aagatatgaa ttcttaatta acgataaaat
     2641 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     2701 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     2761 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     2821 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     2881 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2941 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     3001 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     3061 atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     3121 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     3181 ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg
     3241 aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc
     3301 tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc
     3361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5461 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5521 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5581 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5641 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5701 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5761 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5821 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5881 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5941 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6001 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6061 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6121 acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc
     6181 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     6241 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     6301 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     6361 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     6421 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     6481 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     6541 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     6601 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     6661 tcctccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg
     6721 catccgaatc gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca
     6781 cctcggggg
//
