LOCUS       sequence-11358-e        7054 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 7054)
  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 7054)
  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: 7TP
FEATURES             Location/Qualifiers
     source          1..7054
                     /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..896
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     793..813
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    963..1551
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1016..1036)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1434..1445)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1638..1871
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1943..2064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1980..1999)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2034..2053
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2083..2218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2250..2269)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2251..2269)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2279..2296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2279..2295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2288..2310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2440..2895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2527..2546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2737..2758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2924..3028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3029..3097,3098..3889)
                     /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(3247..3266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4060..4648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4549..4568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4802..4819
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4936..4957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4972..4977,4978..4995,4996..5002)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5010..5026
                     /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     5015..5037
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5034..5050
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5034..5050
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5069..5089
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5071..5089
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             5345..5525
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5569..5694
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6187..6420
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6605..6649
                     /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    6916..7033
                     /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 accatgggta
      301 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
      361 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
      421 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
      481 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
      541 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
      601 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
      661 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
      721 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
      781 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
      841 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
      901 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgccagtcg
      961 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1021 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     1081 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     1141 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     1201 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     1261 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1321 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     1381 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     1441 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     1501 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc
     1561 gagctcggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     1621 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     1681 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     1741 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     1801 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     1861 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     1921 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     1981 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2041 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     2101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2221 ggcttttgcg tcgagacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     2281 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2341 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2401 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat
     2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2641 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2761 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2881 taacgtttac aatttcccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2941 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3001 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3061 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3121 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3181 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3241 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3301 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3361 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3421 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3481 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3541 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3601 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3661 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3721 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3781 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3841 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3901 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3961 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4201 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4261 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4321 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4381 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4441 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4501 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4561 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4621 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4681 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4741 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4801 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4861 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4921 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4981 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5041 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     5101 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     5161 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     5221 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     5281 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacaat
     5341 aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5401 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5461 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     5521 tagcagtggc gcccgaacag ggacctgaaa gcgaaaggga aaccagagct ctctcgacgc
     5581 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     5641 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     5701 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     5761 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     5821 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     5881 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     5941 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     6001 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     6061 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     6121 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     6181 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     6241 ctcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     6301 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     6361 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     6421 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     6481 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     6541 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     6601 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     6661 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     6721 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     6781 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     6841 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     6901 acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     6961 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     7021 aattcaaaat tttatcgatc acgagactag cctc
//
