LOCUS       sequence-267426-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MLV-based transfer vector expressing GFP and neomycin resistance 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  AUTHORS   Kim K, Dauphin A, Komurlu S, McCauley SM, Yurkovetskiy L, Carbone C,
            Diehl WE, Strambio-De-Castillia C, Campbell EM, Luban J
  TITLE     Cyclophilin A protects HIV-1 from restriction by human TRIM5alpha.
  JOURNAL   Nat Microbiol. 2019 Dec;4(12):2044-2051. doi: 
            10.1038/s41564-019-0592-5. Epub 2019 Oct 21.
  PUBMED    31636416
REFERENCE   2  (bases 1 to 6823)
  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/s41564-019-0592-5"; journalName: "Nat Microbiol"; date: 
            "2019-12"; volume: "4"; issue: "12"; pages: "2044-2051"
COMMENT     Sequence Label: pLXIN GFP
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    63..262
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             463..879
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     809..831
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     847..863
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(902..904,905..907,908..1621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(947..968)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1208..1227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1555..1576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1668..2241
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1691..2243
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1835..1852)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2062..2081
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2243..3046
                     /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(2306..2325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2916..2935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3106..3699
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3802..3824)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3910..4050
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3965..3984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4065..4082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4236..4824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4316..4335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4995..5786,5787..5855))
                     /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     5618..5637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5856..5960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6028..6046
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             6648..6823
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
ORIGIN
        1 tgggggctcg tccgggattt ggagacccct gcccagggac caccgaccca ccaccgggag
       61 gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt
      121 atgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      181 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
      241 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
      301 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
      361 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
      421 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
      481 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
      541 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
      601 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
      661 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
      721 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
      781 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
      841 tcctcccttt atccagccct cactccttct ctaggcgccg gaattcgtta acgtcgccac
      901 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1381 cggcatcaag gccaacttca agacccgcca caacatcgag gacggcggcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1621 aggatccact agtaacggcc gccagaattc cgcccctctc cccccccccc ntctccctcc
     1681 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1741 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1801 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1861 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1921 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1981 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2041 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2101 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2161 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2221 ctttgaaaaa cacgatgata atatgggatc ggccattgaa caagatggat tgcacgcagg
     2281 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2341 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2401 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2461 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2521 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2581 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2641 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2701 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2761 gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga cccatggcga
     2821 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2881 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2941 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3001 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3061 ttcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3121 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     3181 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     3241 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3301 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3361 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3421 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3481 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3541 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3601 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3661 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
     3721 gagacccctg cccagggacc accgacccac caccgggagg taagctggct gcctcgcgcg
     3781 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3841 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3901 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     3961 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4021 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4861 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4921 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4981 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5041 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5101 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5161 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5221 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5281 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     5341 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5401 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5461 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5521 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5581 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     5641 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5701 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5761 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5821 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5881 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5941 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6001 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     6061 gaattgctag caattgctag caattgctag caattcatac cagatcaccg aaaactgtcc
     6121 tccaaatgtg tccccctcac actcccaaat tcgcgggctt ctgcctctta gaccactcta
     6181 ccctattccc cacactcacc ggagccaaag ccgcggccct tccgtttctt tgcttttgaa
     6241 agaccccacc cgtaggtggc aagctagctt aagtaacgcc actttgcaag gcatggaaaa
     6301 atacataact gagaatagaa aagttcagat caaggtcagg aacaaagaaa cagctgaata
     6361 ccaaacagga tatctgtggt aagcggttcc tgccccggct cagggccaag aacagatgag
     6421 acagctgagt gatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcggg
     6481 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag tgaatcatca
     6541 gatgtttcca gggtgcccca aggacctgaa aatgaccctg taccttattt gaactaacca
     6601 atcagttcgc ttctcgcttc tgttcgcgcg cttccgctct ccgagctcaa taaaagagcc
     6661 cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat
     6721 tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg
     6781 tctcctctga gtgattgact acccacgacg ggggtctttc att
//
