LOCUS       sequence-190957-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP expressing bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  TITLE     pLXRN bicistronic retroviral vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6812)
  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            "Unpublished"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: pLXRN-EGFP
FEATURES             Location/Qualifiers
     source          1..6812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..184
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(242..814,815..817,818..826)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(407..424)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     634..653
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             849..1652
                     /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(912..931)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1522..1541
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             1712..2305
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2408..2430)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2516..2656
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2571..2590
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2671..2688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3601..4392,4393..4461))
                     /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     4224..4243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4462..4566)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4634..4652
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5254..5429
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5492..5691
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5892..6308
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6238..6260
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6276..6292
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      6316..6325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6322..6324,6325..6327,6328..6812,1..229)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6322..6324,6325..6327,6328..6812,1..226)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6367..6388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6628..6647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
       61 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
      121 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
      181 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac tcgaggatcc
      241 gcccctctcc ctcccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
      301 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
      361 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
      421 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
      481 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
      541 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
      601 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
      661 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
      721 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
      781 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccggg ataattcctg
      841 cagccaatat gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt
      901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
      961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1021 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     1081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     1141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     1201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1381 aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga
     1441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     1501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     1561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     1621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg ataaaataaa
     1681 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     1741 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     1801 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     1861 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     1921 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     1981 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     2041 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     2101 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     2161 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     2221 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     2281 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
     2341 gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg
     2401 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2461 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     2521 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     2581 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2641 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3901 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4201 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4621 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tgctagcaat
     4681 tgctagcaat tgctagcaat tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc
     4741 cctcacactc ccaaattcgc gggcttctgc ctcttagacc actctaccct attccccaca
     4801 ctcaccggag ccaaagccgc ggcccttccg tttctttgct tttgaaagac cccacccgta
     4861 ggtggcaagc tagcttaagt aacgccactt tgcaaggcat ggaaaaatac ataactgaga
     4921 atagaaaagt tcagatcaag gtcaggaaca aagaaacagc tgaataccaa acaggatatc
     4981 tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca gatgagacag ctgagtgatg
     5041 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcggggcca agaacagatg
     5101 gtccccagat gcggtccagc cctcagcagt ttctagtgaa tcatcagatg tttccagggt
     5161 gccccaagga cctgaaaatg accctgtacc ttatttgaac taaccaatca gttcgcttct
     5221 cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa agagcccaca acccctcact
     5281 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
     5341 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     5401 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
     5461 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
     5521 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
     5581 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     5641 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     5701 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     5761 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     5821 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     5881 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     5941 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     6001 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     6061 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     6121 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     6181 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     6241 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     6301 taggcgccgg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     6361 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     6421 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     6481 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     6541 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     6601 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     6661 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     6721 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     6781 atggccgaca agcagaagaa cggcatcaag gt
//
