LOCUS       sequence-192441-        6736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RFP expressing bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6736)
  TITLE     pLXIN2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6736)
  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            "Unpublished"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: pLXIN2-RFP
FEATURES             Location/Qualifiers
     source          1..6736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..28
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             630..805
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    868..1067
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1268..1684
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1614..1636
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1652..1668
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1692..1701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1698..2375
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1878..1898)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2096..2115
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2290..2313
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2388..2959
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2409..2961
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2553..2570)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2780..2799
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2961..3764
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3024..3043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3634..3653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3824..4417
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4520..4542)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4628..4768
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4683..4702
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4783..4800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4954..5542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5034..5053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5713..6504,6505..6573))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6336..6355
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6574..6678)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattgct agcaattgct
       61 agcaattgct agcaattcat accagatcac cgaaaactgt cctccaaatg tgtccccctc
      121 acactcccaa attcgcgggc ttctgcctct tagaccactc taccctattc cccacactca
      181 ccggagccaa agccgcggcc cttccgtttc tttgcttttg aaagacccca cccgtaggtg
      241 gcaagctagc ttaagtaacg ccactttgca aggcatggaa aaatacataa ctgagaatag
      301 aaaagttcag atcaaggtca ggaacaaaga aacagctgaa taccaaacag gatatctgtg
      361 gtaagcggtt cctgccccgg ctcagggcca agaacagatg agacagctga gtgatgggcc
      421 aaacaggata tctgtggtaa gcagttcctg ccccggctcg gggccaagaa cagatggtcc
      481 ccagatgcgg tccagccctc agcagtttct agtgaatcat cagatgtttc cagggtgccc
      541 caaggacctg aaaatgaccc tgtaccttat ttgaactaac caatcagttc gcttctcgct
      601 tctgttcgcg cgcttccgct ctccgagctc aataaaagag cccacaaccc ctcactcggc
      661 gcgccagtct tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg
      721 ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
      781 ctacccacga cgggggtctt tcatttgggg gctcgtccgg gatttggaga cccctgccca
      841 gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga
      901 ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg tactagttag ctaactagct
      961 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
     1021 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
     1081 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     1141 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     1201 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1261 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1321 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1381 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1441 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1501 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1561 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1621 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1681 cgccggaatt cgccaccatg gcctcctccg agaacgtcat caaggagttc atgcgcttca
     1741 aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1801 gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc ggccccctgc
     1861 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc
     1921 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     1981 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2041 acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg
     2101 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     2161 gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa ggacggcggc cactacctgg
     2221 tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg
     2281 tggactccaa gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg
     2341 agcgcaccga gggccgccac cacctgttcc tgtagctcga ggatccgccc ctctccctcc
     2401 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     2461 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     2521 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     2581 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaagcaa cgtctgtagc
     2641 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     2701 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagtaggat
     2761 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     2821 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     2881 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     2941 ttgaaaaaca cgatgataat atgggatcgg ccattgaaca agatggattg cacgcaggtt
     3001 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3061 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3121 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3181 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3241 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3301 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3361 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3421 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3481 tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
     3541 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3601 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3661 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3721 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3781 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     3841 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     3901 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     3961 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     4021 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4081 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     4141 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4201 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4261 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4321 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4381 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     4441 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     4501 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4561 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4621 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     4681 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4741 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     4801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5881 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6001 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     6061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6241 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6301 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     6361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6421 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6541 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6661 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6721 tattatcatg acatta
//
