LOCUS       sequence-164348-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing dKatushka2 fluorescent protein driven 
            by SFFV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  AUTHORS   Mohme M, Maire CL, Riecken K, Zapf S, Aranyossy T, Westphal M, 
            Lamszus K, Fehse B
  TITLE     Optical Barcoding for Single-Clone Tracking to Study Tumor 
            Heterogeneity.
  JOURNAL   Mol Ther. 2017 Jan 18. pii: S1525-0016(16)45496-1. doi: 
            10.1016/j.ymthe.2016.12.014.
  PUBMED    28109958
REFERENCE   2  (bases 1 to 7132)
  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. 
            2017 Jan 18. pii: S1525-0016(16)45496-1. doi: 
            10.1016/j.ymthe.2016.12.014."
COMMENT     Sequence Label: LeGO-dKatushka2
FEATURES             Location/Qualifiers
     source          1..7132
                     /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    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2620..2636
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2678..2711)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2778..3184
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3190..3199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3932..3965)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4021..4609
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4074..4094)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4492..4503)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4612..4628)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5138..5318
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5380..5968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5460..5479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6139..6930,6931..6999))
                     /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     6762..6781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7000..7104)
                     /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 gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctagtc gaggtcgacg gtatcgataa
     2641 gctcgcttca cgagattcca gcaggtcgag ggacctaata acttcgtata gcatacatta
     2701 tacgaagtta tattaagggt tccaagctta agcggccgct gaaagacccc acctgtaggt
     2761 ttggcaagct agctgcagta acgccatttt gcaaggcatg gaaaaatacc aaaccaagaa
     2821 tagagaagtt cagatcaagg gcgggtacat gaaaatagct aacgttgggc caaacaggat
     2881 atctgcggtg agcagtttcg gccccggccc ggggccaaga acagatggtc accgcagttt
     2941 cggccccggc ccgaggccaa gaacagatgg tccccagata tggcccaacc ctcagcagtt
     3001 tcttaagacc catcagatgt ttccaggctc ccccaaggac ctgaaatgac cctgcgcctt
     3061 atttgaatta accaatcagc ctgcttctcg cttctgttcg cgcgcttctg cttcccgagc
     3121 tctataaaag agctcacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc
     3181 ccggatcccg ccaccatggt gggcgaggac agcgtgctga tcaccgagaa catgcacatg
     3241 aaactctaca tggagggcac cgtgaacgac caccacttca agtgcacatc cgagggcgaa
     3301 ggcaagccct acgagggcac ccagaccatg aagatcaagg tggtcgaggg cggccctctc
     3361 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac ctttatcaac
     3421 cacacccagg gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag
     3481 aggatcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     3541 aacggctgcc tcatctacaa cgtcaagatc aacggggtga acttcccatc caacggccct
     3601 gtgatgcaga agaaaacact cggctgggag gccagcaccg agatgctgta ccccgctgac
     3661 agcggcctga gaggccatgc ccagatggcc ctgaagctcg tgggcggggg ctacctgcac
     3721 tgctccctca agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
     3781 ttctacttcg tggacaggag actggaaaga atcaaggagg ccgacaaaga gacctacgtc
     3841 gagcagcacg agatggctgt ggccaggtac tgcgacctgc ctagcaaact ggggcacagc
     3901 ctgtacaagt aagaattcgt cgagggacct aataacttcg tatagcatac attatacgaa
     3961 gttatacatg tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat
     4021 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4081 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4141 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4201 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4261 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4321 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4381 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4441 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4501 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4561 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     4621 cgacctcgat cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     4681 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     4741 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     4801 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     4861 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
     4921 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
     4981 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
     5041 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
     5101 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     5161 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5221 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5281 cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca
     5341 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5401 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5461 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5521 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5581 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5641 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5701 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5761 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5821 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5881 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5941 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6001 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6061 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6121 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6181 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6241 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6301 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6361 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6421 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6481 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6541 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6601 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6661 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6721 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6781 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6841 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6901 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6961 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7081 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
