LOCUS       sequence-209079-        8537 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 8537)
  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 8537)
  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-iT2
FEATURES             Location/Qualifiers
     source          1..8537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2619..2635
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2677..2710)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2781..3187
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    join(3272..3846,3847..3849,3850..3858)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3439..3456)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3666..3685
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3859..5289
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(5336..5369)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5425..6013
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5478..5498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5896..5907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6016..6032)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6542..6722
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6784..7372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6864..6883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7543..8334,8335..8403))
                     /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     8166..8185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8404..8508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagtcg aggtcgacgg tatcgataag
     2641 ctcgcttcac gagattccag caggtcgagg gacctaataa cttcgtatag catacattat
     2701 acgaagttat attaagggtt ccaagcttaa gcggccggcc gctgaaagac cccacctgta
     2761 ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat accaaaccaa
     2821 gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg ggccaaacag
     2881 gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg gtcaccgcag
     2941 tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca accctcagca
     3001 gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat gaccctgcgc
     3061 cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg
     3121 agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga ttgactgagt
     3181 cgcccggatc ccagtgtggt ggtacgggaa ttcctgcagg cctcgacgag ggccggcgcg
     3241 ccgcggccgc tacgtaaatt ccgccccccc ccccctctcc ctcccccccc cctaacgtta
     3301 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     3361 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     3421 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     3481 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     3541 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     3601 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     3661 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     3721 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     3781 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     3841 gataatatgg ccacaaccat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
     3901 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     3961 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     4021 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     4081 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     4141 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     4201 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     4261 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     4321 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     4381 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     4441 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     4501 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc
     4561 agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat caaagagttc
     4621 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     4681 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     4741 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg
     4801 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc
     4861 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc
     4921 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     4981 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     5041 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
     5101 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     5161 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     5221 gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg
     5281 tacaagtaaa gcggccggcc gccagcacag tggtcgaaat tcgtcgaggg acctaataac
     5341 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
     5401 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     5461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5821 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6001 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     6061 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     6121 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     6181 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6241 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     6301 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     6361 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     6421 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     6481 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     6541 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6601 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6661 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6721 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7201 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgacg
//
