LOCUS       sequence-192204-        6777 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 6777)
  TITLE     pLXIN2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6777)
  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            "Unpublished"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-EGFP
FEATURES             Location/Qualifiers
     source          1..6777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    223..794
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    244..796
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(388..405)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     615..634
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             796..1599
                     /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(859..878)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1469..1488
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             1659..2252
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2355..2377)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2463..2603
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2518..2537
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2618..2635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2789..3377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2869..2888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3548..4339,4340..4408))
                     /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     4171..4190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4409..4513)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4581..4599
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5201..5376
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5439..5638
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5839..6255
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6185..6207
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6223..6239
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(6268..6270,6271..6273,6274..6777,1..210)
                     /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(6268..6270,6271..6273,6274..6777,1..207)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6313..6334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6574..6593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
       61 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
      121 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
      181 actctcggca tggacgagct gtacaagtaa ctcgaggatc cgcccctctc cctccccccc
      241 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
      301 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
      361 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
      421 tcgtgaagga agcagttcct ctggaagctt cttgaagaca agcaacgtct gtagcgaccc
      481 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
      541 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt aggatagttg
      601 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
      661 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
      721 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
      781 aaacacgatg ataatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
      841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      961 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     1021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     1081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     1141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     1201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     1261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     1321 aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc
     1381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     1441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     1501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     1561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgata
     1621 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     1681 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
     1741 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
     1801 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     1861 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     1921 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     1981 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     2041 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     2101 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     2161 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     2221 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
     2281 ctgcccaggg accaccgacc caccaccggg aggtaagctg gctgcctcgc gcgtttcggt
     2341 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2401 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2461 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2521 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2581 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3841 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4141 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     4201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4501 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4561 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattgc
     4621 tagcaattgc tagcaattgc tagcaattca taccagatca ccgaaaactg tcctccaaat
     4681 gtgtccccct cacactccca aattcgcggg cttctgcctc ttagaccact ctaccctatt
     4741 ccccacactc accggagcca aagccgcggc ccttccgttt ctttgctttt gaaagacccc
     4801 acccgtaggt ggcaagctag cttaagtaac gccactttgc aaggcatgga aaaatacata
     4861 actgagaata gaaaagttca gatcaaggtc aggaacaaag aaacagctga ataccaaaca
     4921 ggatatctgt ggtaagcggt tcctgccccg gctcagggcc aagaacagat gagacagctg
     4981 agtgatgggc caaacaggat atctgtggta agcagttcct gccccggctc ggggccaaga
     5041 acagatggtc cccagatgcg gtccagccct cagcagtttc tagtgaatca tcagatgttt
     5101 ccagggtgcc ccaaggacct gaaaatgacc ctgtacctta tttgaactaa ccaatcagtt
     5161 cgcttctcgc ttctgttcgc gcgcttccgc tctccgagct caataaaaga gcccacaacc
     5221 cctcactcgg cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg tattcccaat
     5281 aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc
     5341 tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag
     5401 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
     5461 tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta
     5521 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
     5581 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
     5641 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
     5701 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
     5761 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     5821 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     5881 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     5941 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
     6001 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
     6061 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
     6121 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     6181 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     6241 ccttctctag gcgccggaat tcgcaccatg gtgagcaagg gcgaggagct gttcaccggg
     6301 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     6361 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     6421 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     6481 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     6541 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     6601 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     6661 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     6721 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccac
//
