LOCUS       sequence-176311-        7793 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Weber K, Bartsch U, Stocking C, Fehse B
  TITLE     A multicolor panel of novel lentiviral ""gene ontology"" (LeGO) 
            vectors for functional gene analysis.
  JOURNAL   Mol Ther. 2008 Apr . 16(4):698-706.
  PUBMED    18362927
REFERENCE   2  (bases 1 to 7793)
  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: "Mol Ther. 
            2008 Apr . 16(4):698-706."
COMMENT     Sequence Label: LeGO-iG2
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    14..566
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(158..175)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     385..404
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(578..580,581..583,584..1297)
                     /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(578..580,581..583,584..1294)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(623..644)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(884..903)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1231..1252
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1344..1377)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1433..2021
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1486..1506)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1904..1915)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2024..2040)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2550..2730
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2792..3380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2872..2891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3551..4342,4343..4411))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4174..4193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4412..4516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4591..4610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4782..5161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5163..5361
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5316..5336
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5379..5559
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5606..5731
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6228..6461
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6646..6690
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6988..7105
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     7158..7174
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7216..7249)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7320..7726
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
       61 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
      121 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
      181 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
      241 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
      301 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
      361 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
      421 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
      481 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
      541 ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct
      601 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      661 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      721 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
      781 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      841 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      901 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      961 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1021 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1081 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1141 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1201 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1261 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccggccgc cagcacagtg
     1321 gtcgaaattc gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca
     1381 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     1441 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1501 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1561 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1621 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1681 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1741 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1801 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     1861 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1921 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     1981 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     2041 atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     2101 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     2161 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     2221 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     2281 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     2341 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     2401 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     2461 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     2521 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     2581 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     2641 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     2701 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
     2761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     4561 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4621 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     4681 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     4741 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     4801 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4861 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4921 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4981 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5041 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5101 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     5161 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     5221 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     5281 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     5341 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     5401 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5461 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5521 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     5581 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     5641 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     5701 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     5761 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     5821 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     5881 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     5941 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     6001 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     6061 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     6121 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     6181 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     6241 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     6301 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     6361 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     6421 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     6481 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     6541 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     6601 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     6661 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     6721 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     6781 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     6841 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     6901 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     6961 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     7021 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     7081 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     7141 gttagtaccg ggcccgccga ggtcgacggt atcgataagc tcgcttcacg agattccagc
     7201 aggtcgaggg acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc
     7261 caagcttaag cggccggccg ctgaaagacc ccacctgtag gtttggcaag ctagctgcag
     7321 taacgccatt ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa
     7381 gggcgggtac atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt
     7441 cggccccggc ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc
     7501 aagaacagat ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat
     7561 gtttccaggc tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca
     7621 gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca
     7681 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccggatcc cagtgtggtg
     7741 gtacgggaat tcctgcaggc ctcgacgagg gccggcgcgc cgcggccgct acg
//
