LOCUS       sequence-12900-e        7856 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 7856)
  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 7856)
  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-1xT/BSD
FEATURES             Location/Qualifiers
     source          1..7856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..2935
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2854..2874
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2952..2968
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3010..3043)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3110..3516
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3522..3531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3528..4229
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3661..3679)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3929..3948
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4123..4146
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4230..4625
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(4656..4689)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4745..5333
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4798..4818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5216..5227)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5336..5352)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5862..6042
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6104..6692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6184..6203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6863..7654,7655..7723))
                     /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     7486..7505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7724..7828)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg ccgccatctc
     2641 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2701 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2761 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2821 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2881 gtaaagtaat tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgttaacgc
     2941 gcggtgaccc tcgaggtcga cggtatcgat aagctcgctt cacgagattc cagcaggtcg
     3001 agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttccaagct
     3061 taagcggccg ctgaaagacc ccacctgtag gtttggcaag ctagctgcag taacgccatt
     3121 ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac
     3181 atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc
     3241 ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat
     3301 ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc
     3361 tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct
     3421 cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact
     3481 cggcgcgcca gtcctccgat tgactgagtc gcccggatcc cgccaccatg gtgagcaagg
     3541 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     3601 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     3661 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     3721 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     3781 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     3841 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     3901 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3961 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     4021 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     4081 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     4141 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     4201 tcctgtacgg catggacgag ctgtacaagg ccaagccttt gtctcaagaa gaatccaccc
     4261 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     4321 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     4381 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     4441 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     4501 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     4561 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4621 gctaatgtac aagtaagaat tcgtcgaggg acctaataac ttcgtatagc atacattata
     4681 cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat
     4741 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4801 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4861 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4921 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4981 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5041 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5101 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5161 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5221 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5281 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     5341 ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     5401 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     5461 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5521 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     5581 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
     5641 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     5701 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     5761 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     5821 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     5881 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5941 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6001 gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc
     6061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6541 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6841 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6901 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6961 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7021 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7081 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7141 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7201 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7261 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7321 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7381 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7441 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7501 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7561 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7621 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7681 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7741 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7801 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
