LOCUS       sequence-11355-e        8051 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 8051)
  AUTHORS   Fuerer C, Nusse R
  TITLE     Lentiviral vectors to probe and manipulate the wnt signaling 
            pathway.
  JOURNAL   PLoS One. 2010  . 5(2):e9370.
  PUBMED    20186325
REFERENCE   2  (bases 1 to 8051)
  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: "PLoS One. 
            2010  . 5(2):e9370."
COMMENT     Sequence Label: 7TF
FEATURES             Location/Qualifiers
     source          1..8051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        201..232
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      288..297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             294..1946
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(360..378)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    1960..2548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2013..2033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2431..2442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2635..2868
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2940..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2977..2996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3031..3050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3080..3215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3247..3266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3248..3266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3276..3293)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3276..3292)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3285..3307)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3437..3892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3524..3543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3734..3755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3921..4025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4026..4094,4095..4886)
                     /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(4244..4263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5057..5645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5546..5565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5799..5816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5933..5954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5969..5974,5975..5992,5993..5999)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6007..6023
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6012..6034
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6031..6047
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6031..6047
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6066..6086
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6068..6086
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             6342..6522
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6566..6691
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7184..7417
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7602..7646
                     /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    7913..8030
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctagcccggg ctcgagcagc tgaagcttgc atgcctgcag ggtaccgagc tcttacgcga
       61 gatcaaaggg ggtaagatca aagggggtaa gatcaaaggg gcgcgagatc aaagggggta
      121 agatcaaagg gggtaagatc aaagggggta agatcaaagg ggcgcgcccg cgtgctagcc
      181 cgggctcgag atctagactc tagagggtat ataatggaag ctcgaattcc agcttggcat
      241 tccggtactg ttggtaaaaa gcttggcatt ccggtactgt tggtaaagcc accatggaag
      301 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
      361 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
      421 cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt
      481 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
      541 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
      601 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc
      661 ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc
      721 tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
      781 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc
      841 cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg
      901 gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca
      961 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1021 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     1081 tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc
     1141 aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca
     1201 aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag
     1261 tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca
     1321 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     1381 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     1441 gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa
     1501 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     1561 tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta
     1621 agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca
     1681 acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     1741 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     1801 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     1861 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     1921 agaagggcgg aaagatcgcc gtgtaattct agagtcgaca atcaacctct ggattacaaa
     1981 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2041 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2101 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2161 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2221 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2281 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2341 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2401 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2461 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2521 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct ttaagaccaa
     2581 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     2641 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     2701 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2761 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2821 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     2881 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     2941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3061 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     3121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcgtcg agacgtaccc
     3241 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     3301 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3361 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3421 aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3721 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3781 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3841 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcccaggtg
     3901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4261 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4561 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5761 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5821 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5881 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5941 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6001 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     6061 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     6121 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     6181 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     6241 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     6301 cagagatatt gtatttaagt gcctagctcg atacaataaa cgggtctctc tggttagacc
     6361 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6421 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6481 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     6541 cctgaaagcg aaagggaaac cagagctctc tcgacgcagg actcggcttg ctgaagcgcg
     6601 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     6661 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6721 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6781 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     6841 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     6901 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     6961 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7021 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7081 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7141 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7201 ggttcttggg agcagcagga agcactatgg gcgcagcctc aatgacgctg acggtacagg
     7261 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7321 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7381 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7441 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7501 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7561 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7621 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7681 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7741 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7801 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7861 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa
     7921 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     7981 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg
     8041 agactagcct c
//
