LOCUS       sequence-11364-e        7102 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 7102)
  AUTHORS   ten Berge D, Koole W, Fuerer C, Fish M, Eroglu E, Nusse R
  TITLE     Wnt signaling mediates self-organization and axis formation in 
            embryoid bodies.
  JOURNAL   Cell Stem Cell. 2008 Nov 6. 3(5):508-18.
  PUBMED    18983966
REFERENCE   2  (bases 1 to 7102)
  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: "Cell Stem 
            Cell. 2008 Nov 6. 3(5):508-18."
COMMENT     Sequence Label: 7TC
FEATURES             Location/Qualifiers
     source          1..7102
                     /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..1004
                     /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(439..457)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     707..726
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1011..1599
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1064..1084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1482..1493)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1686..1919
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1991..2112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2028..2047)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2082..2101
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2131..2266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2298..2317)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2299..2317)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2327..2344)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2327..2343)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2336..2358)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2488..2943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2575..2594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2785..2806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2972..3076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3077..3145,3146..3937)
                     /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(3295..3314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4108..4696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4597..4616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4850..4867
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4984..5005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5020..5025,5026..5043,5044..5050)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5058..5074
                     /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     5063..5085
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5082..5098
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5082..5098
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5117..5137
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5119..5137
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             5393..5573
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5617..5742
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6235..6468
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6653..6697
                     /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    6964..7081
                     /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 accatggtga
      301 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      361 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      421 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      481 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      541 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      601 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
      661 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
      721 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
      781 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
      841 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
      901 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
      961 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagtcgac aatcaacctc
     1021 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1081 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1141 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     1201 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     1261 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     1321 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1381 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     1441 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     1501 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     1561 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacc
     1621 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     1681 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     1741 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     1801 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     1861 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     1921 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     1981 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2041 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2101 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc
     2161 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     2221 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcgtc
     2281 gagacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     2341 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2401 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2461 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
     2521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2941 tttcccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3001 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3061 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3121 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3181 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3241 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3301 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3361 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3421 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3481 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3541 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3601 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3661 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3721 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3781 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3841 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3901 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3961 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4021 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4081 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4141 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4201 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4261 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4321 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4381 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4441 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4501 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4561 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4621 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4681 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4741 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4801 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4861 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4921 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4981 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5041 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5101 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     5161 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
     5221 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
     5281 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
     5341 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacaataa acgggtctct
     5401 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     5461 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5521 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     5581 ccgaacaggg acctgaaagc gaaagggaaa ccagagctct ctcgacgcag gactcggctt
     5641 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     5701 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     5761 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     5821 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     5881 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     5941 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     6001 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     6061 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     6121 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     6181 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     6241 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcct caatgacgct
     6301 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     6361 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     6421 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     6481 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     6541 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     6601 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     6661 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     6721 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     6781 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     6841 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     6901 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     6961 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     7021 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     7081 tatcgatcac gagactagcc tc
//
