LOCUS       sequence-192445-        6057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bicistronic retroviral vector (optimized internal ribosome entry 
            site).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6057)
  TITLE     pLXIN2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6057)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..6057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    227..426
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             627..1043
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     973..995
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1011..1027
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1068..1639
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1089..1641
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1233..1250)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1460..1479
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1641..2444
                     /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(1704..1723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2314..2333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             2504..3097
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3200..3222)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3308..3448
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3363..3382
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3463..3480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3634..4222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3714..3733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4393..5184,5185..5253))
                     /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     5016..5035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5254..5358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5426..5444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             join(6046..6057,1..164)
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
ORIGIN
        1 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
       61 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      121 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      181 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      241 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      301 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      361 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      421 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      481 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      541 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      601 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
      661 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
      721 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
      781 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
      841 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
      901 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
      961 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1021 ccctcactcc ttctctaggc gccggaattc gttaactcga ggatccgccc ctctccctcc
     1081 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1141 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1201 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1261 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaagcaa cgtctgtagc
     1321 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     1381 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagtaggat
     1441 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     1501 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     1561 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     1621 ttgaaaaaca cgatgataat atgggatcgg ccattgaaca agatggattg cacgcaggtt
     1681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     1861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2161 tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
     2221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     2461 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     2521 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     2581 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     2641 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     2701 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     2761 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     2821 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     2881 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     2941 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     3001 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     3061 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     3121 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     3181 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3241 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3301 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     3361 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     3421 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     3481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4681 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4981 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     5041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5401 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     5461 attgctagca attgctagca attgctagca attcatacca gatcaccgaa aactgtcctc
     5521 caaatgtgtc cccctcacac tcccaaattc gcgggcttct gcctcttaga ccactctacc
     5581 ctattcccca cactcaccgg agccaaagcc gcggcccttc cgtttctttg cttttgaaag
     5641 accccacccg taggtggcaa gctagcttaa gtaacgccac tttgcaaggc atggaaaaat
     5701 acataactga gaatagaaaa gttcagatca aggtcaggaa caaagaaaca gctgaatacc
     5761 aaacaggata tctgtggtaa gcggttcctg ccccggctca gggccaagaa cagatgagac
     5821 agctgagtga tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcggggc
     5881 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagtg aatcatcaga
     5941 tgtttccagg gtgccccaag gacctgaaaa tgaccctgta ccttatttga actaaccaat
     6001 cagttcgctt ctcgcttctg ttcgcgcgct tccgctctcc gagctcaata aaagagc
//
