LOCUS       sequence-204009-        7135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7135)
  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 7135)
  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-V2
FEATURES             Location/Qualifiers
     source          1..7135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     607..627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             670..850
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    897..1022
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1519..1752
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1937..1981
                     /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    2279..2396
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2455..2471
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2513..2546)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2613..3019
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3025..3034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3031..3033,3034..3036,3037..3750)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3076..3097)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3337..3356)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3684..3705
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3770..3803)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3859..4447
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3912..3932)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4330..4341)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4450..4466)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             4976..5156
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5218..5806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5298..5317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5977..6768,6769..6837))
                     /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     6600..6619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6838..6942)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7017..7036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
       61 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      121 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      181 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      241 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      301 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      361 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      421 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      481 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      541 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      601 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt
      661 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      721 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      781 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
      841 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
      901 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      961 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     1021 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     1081 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     1141 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     1201 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     1261 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     1321 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag
     1381 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     1441 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     1501 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     1561 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     1621 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     1681 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     1741 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     1801 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     1861 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     1921 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     1981 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     2041 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     2101 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     2161 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     2221 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
     2281 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     2341 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     2401 tttattacag ggacagcaga gatccagttt ggttagtacc gggcccgctc tagtcgaggt
     2461 cgacggtatc gataagctcg cttcacgaga ttccagcagg tcgagggacc taataacttc
     2521 gtatagcata cattatacga agttatatta agggttccaa gcttaagcgg ccgctgaaag
     2581 accccacctg taggtttggc aagctagctg cagtaacgcc attttgcaag gcatggaaaa
     2641 ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa tagctaacgt
     2701 tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc caagaacaga
     2761 tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc agatatggcc
     2821 caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca aggacctgaa
     2881 atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct gttcgcgcgc
     2941 ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg ccagtcctcc
     3001 gattgactga gtcgcccgga tcccgccacc atggtgagca agggcgagga gctgttcacc
     3061 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3121 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     3181 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag
     3241 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3301 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3361 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3421 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3481 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     3541 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3601 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     3661 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3721 actctcggca tggacgagct gtacaagtaa gaattcgtcg agggacctaa taacttcgta
     3781 tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta ggtacaattc
     3841 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3901 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3961 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4021 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4081 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4141 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4201 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4261 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4321 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4381 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4441 tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag
     4501 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     4561 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     4621 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4681 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     4741 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     4801 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     4861 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     4921 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     4981 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5041 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5101 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat
     5161 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5221 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5281 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5341 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5401 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5461 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5521 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5581 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5641 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5701 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5761 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5821 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5881 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5941 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6001 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6061 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6121 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6181 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6241 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6301 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6361 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6421 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6481 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6541 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6601 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6661 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6721 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6781 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6841 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6901 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6961 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7021 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7081 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgaca
//
