LOCUS       sequence-354475-        7134 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 7134)
  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 7134)
  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-Cer2
FEATURES             Location/Qualifiers
     source          1..7134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(515..921)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     protein_bind    988..1021
                     /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(1063..1079)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1137..1254
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1552..1596)
                     /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    1781..2014
                     /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    2511..2636
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2683..2863
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2881..3079)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2906..2926)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3081..3460
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3632..3651
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3726..3830
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3831..3899,3900..4691)
                     /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(4049..4068)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4862..5450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5351..5370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5512..5692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     6202..6218
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6221..6809
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6327..6338
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6736..6756
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    6865..6898
                     /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(6918..7134,1..497,498..500,501..503))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6963..6984)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtcg
       61 ctgatggcgt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
      301 ccccaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catggtggcg ggatccgggc gactcagtca atcggaggac
      541 tggcgcgccg agtgaggggt tgtgagctct tttatagagc tcgggaagca gaagcgcgcg
      601 aacagaagcg agaagcaggc tgattggtta attcaaataa ggcgcagggt catttcaggt
      661 ccttggggga gcctggaaac atctgatggg tcttaagaaa ctgctgaggg ttgggccata
      721 tctggggacc atctgttctt ggcctcgggc cggggccgaa actgcggtga ccatctgttc
      781 ttggccccgg gccggggccg aaactgctca ccgcagatat cctgtttggc ccaacgttag
      841 ctattttcat gtacccgccc ttgatctgaa cttctctatt cttggtttgg tatttttcca
      901 tgccttgcaa aatggcgtta ctgcagctag cttgccaaac ctacaggtgg ggtctttcag
      961 cggccgctta agcttggaac ccttaatata acttcgtata atgtatgcta tacgaagtta
     1021 ttaggtccct cgacctgctg gaatctcgtg aagcgagctt atcgataccg tcgacctcga
     1081 tagagcgggc ccggtactaa ccaaactgga tctctgctgt ccctgtaata aacccgaaaa
     1141 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
     1201 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
     1261 atgaatactg ccatttgtct gcagaattgg cgcacgcagt gccgatccgt tcactaatcg
     1321 aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg gcctgtcggg
     1381 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
     1441 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
     1501 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
     1561 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
     1621 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
     1681 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
     1741 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
     1801 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
     1861 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
     1921 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
     1981 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
     2041 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
     2101 atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag gtctgaagat
     2161 cagcggccgg ccgcttgctg tgcggtggtc ttacttttgt tttgctcttc ctctatcttg
     2221 tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata gagggttgct
     2281 actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg ttgtagctgt
     2341 cccagtattt gtctacagcc ttctgatgtt tctaacaggc caggattaac tgcgaatcgt
     2401 tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt tccccctggc
     2461 cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact gacgctctcg
     2521 cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact caccagtcgc
     2581 cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc
     2641 tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag attttccaca
     2701 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     2761 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     2821 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag gcaaaacgcg
     2881 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     2941 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     3001 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     3061 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     3121 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     3181 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     3241 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     3301 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     3361 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     3421 gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt
     3481 acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc
     3541 aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca
     3601 caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     3661 caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa
     3721 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3781 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3841 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3901 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3961 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4021 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4081 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4141 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4201 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4261 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4321 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4381 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4441 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4501 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4561 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4621 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4681 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4741 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4801 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4861 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4921 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4981 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5041 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5101 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5161 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5221 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5281 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5341 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5401 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5461 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ctgctagaga
     5521 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     5581 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     5641 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagt
     5701 ccggatgcag ctctcgggcc atgtgatgaa atgctaggcg gctgtcaaac ctccactcta
     5761 atacttctct ctccgggtca tccatcccat gcaggctcac agggtgtaac aagcgggtgt
     5821 tctctccttc attggcttct tctaccttct cttgctcaac tggtactagc ttgtagcacc
     5881 atccaaaggt cagtggatat ctgatccctg gccctggtgt gtagttctgc caatcaggga
     5941 agtagccttg tgtgtggtag atccacagat caaggatatc ttgtcttcgt tgggagtgaa
     6001 ttagcccttc cagtcccccc ttttctttta aaaagtggct aagatctaca gctgccttgt
     6061 aagtcattgg tcttaaaggt acctgaggtg tgactggaaa acccacctcc tcctcctctt
     6121 gtgcttctag ccaggcacaa tcagcattgg tagctgctgt attgctactt gtgattgctc
     6181 catgtttttc taggtctcga tcgaggtcga cggtatcgat gcggggaggc ggcccaaagg
     6241 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     6301 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     6361 ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc
     6421 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     6481 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     6541 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     6601 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     6661 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     6721 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     6781 tcacaaattt tgtaatccag aggttgatta tcgataagct tgatatcgaa ttgtacctag
     6841 tggaaccgga acccttaaac atgtataact tcgtataatg tatgctatac gaagttatta
     6901 ggtccctcga cgaattctta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     6961 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg
     7021 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     7081 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcgg
//
