LOCUS       sequence-354663-        7125 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 7125)
  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 7125)
  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-C2
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..164
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      306..315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(321..727)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     protein_bind    794..827
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(869..885)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    943..1060
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1358..1402)
                     /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    1587..1820
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2317..2442
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2489..2669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2687..2885)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2712..2732)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2887..3266
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3438..3457
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3532..3636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3637..3705,3706..4497)
                     /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     complement(3855..3874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4668..5256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5157..5176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5318..5498
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     6008..6024
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6027..6615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6133..6144
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6542..6562
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    6671..6704
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(6724..7125,1..309))
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7002..7021)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
       61 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
      121 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
      181 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
      241 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
      301 gctcaccatg gtggcgggat ccgggcgact cagtcaatcg gaggactggc gcgccgagtg
      361 aggggttgtg agctctttta tagagctcgg gaagcagaag cgcgcgaaca gaagcgagaa
      421 gcaggctgat tggttaattc aaataaggcg cagggtcatt tcaggtcctt gggggagcct
      481 ggaaacatct gatgggtctt aagaaactgc tgagggttgg gccatatctg gggaccatct
      541 gttcttggcc tcgggccggg gccgaaactg cggtgaccat ctgttcttgg ccccgggccg
      601 gggccgaaac tgctcaccgc agatatcctg tttggcccaa cgttagctat tttcatgtac
      661 ccgcccttga tctgaacttc tctattcttg gtttggtatt tttccatgcc ttgcaaaatg
      721 gcgttactgc agctagcttg ccaaacctac aggtggggtc tttcagcggc cgcttaagct
      781 tggaaccctt aatataactt cgtataatgt atgctatacg aagttattag gtccctcgac
      841 ctgctggaat ctcgtgaagc gagcttatcg ataccgtcga cctcgctaga gcgggcccgg
      901 tactaaccaa actggatctc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
      961 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     1021 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     1081 ttgtctgcag aattggcgca cgcagtgccg atccgttcac taatcgaatg gatctgtctc
     1141 tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg
     1201 ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa
     1261 gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg aataatttta
     1321 tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt
     1381 ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta
     1441 ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa
     1501 tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga
     1561 gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag
     1621 ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt
     1681 gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc
     1741 tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga
     1801 acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct
     1861 ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat ttatataatt
     1921 cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc ggccggccgc
     1981 ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt
     2041 ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa
     2101 tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct
     2161 acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct
     2221 tgcccatact atatgtttta atttatattt tttctttccc cctggcctta accgaatttt
     2281 ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc
     2341 cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct
     2401 tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt
     2461 cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt
     2521 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     2581 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     2641 gctcagatct ggtctaacca gagagaccca gtacaggcaa aacgcgctgc ttatatagac
     2701 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2761 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2821 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2881 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2941 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     3001 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3061 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     3121 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     3181 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     3241 ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag
     3301 cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt
     3361 tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag
     3421 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc
     3481 gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3541 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3601 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3661 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3721 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3781 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3841 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3901 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3961 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4021 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4081 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4141 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4201 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4261 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4321 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4381 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4441 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4501 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4561 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5281 cggttcctgg ccttttgctg gccttttgct cacatgctgc tagagatttt ccacactgac
     5341 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
     5401 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     5461 agctcccagg ctcagatctg gtctaaccag agagacccag tacagtccgg atgcagctct
     5521 cgggccatgt gatgaaatgc taggcggctg tcaaacctcc actctaatac ttctctctcc
     5581 gggtcatcca tcccatgcag gctcacaggg tgtaacaagc gggtgttctc tccttcattg
     5641 gcttcttcta ccttctcttg ctcaactggt actagcttgt agcaccatcc aaaggtcagt
     5701 ggatatctga tccctggccc tggtgtgtag ttctgccaat cagggaagta gccttgtgtg
     5761 tggtagatcc acagatcaag gatatcttgt cttcgttggg agtgaattag cccttccagt
     5821 cccccctttt cttttaaaaa gtggctaaga tctacagctg ccttgtaagt cattggtctt
     5881 aaaggtacct gaggtgtgac tggaaaaccc acctcctcct cctcttgtgc ttctagccag
     5941 gcacaatcag cattggtagc tgctgtattg ctacttgtga ttgctccatg tttttctagg
     6001 tctcgatcga ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt
     6061 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     6121 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     6181 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     6241 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     6301 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     6361 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     6421 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     6481 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     6541 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     6601 atccagaggt tgattatcga taagcttgat atcgaattgt acctagtgga accggaaccc
     6661 ttaaacatgt ataacttcgt ataatgtatg ctatacgaag ttattaggtc cctcgacgaa
     6721 ttcttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
     6781 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
     6841 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
     6901 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
     6961 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
     7021 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     7081 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaa
//
