LOCUS       sequence-200158-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of doubleTAL staple protein dT25 with N-terminally fused 
            cycle3-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  AUTHORS   Praetorius F, Dietz H
  TITLE     Self-assembly of genetically encoded DNA-protein hybrid nanoscale 
            shapes.
  JOURNAL   Science. 2017 Mar 24;355(6331). pii: eaam5488. doi: 
            10.1126/science.aam5488.
  PUBMED    28336611
REFERENCE   2  (bases 1 to 7971)
  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: "Science."; 
            date: "2017-03-24"; volume: "355(6331). pii: eaam5488. doi"; pages: 
            " 10.1126/science.aam5488"
COMMENT     Sequence Label: pSB1A3_GFP_dT25
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(130..149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      943..1531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1432..1451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1685..1702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1726..1783)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     1801..1820
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1801..1819
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1851..1873
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1880..2593
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1910..1938)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2533..2553
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             7601..7618
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             7619..7636
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      7645..7688
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(7692..7710)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7778..7882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7883..7951,7952..7971,1..772)
                     /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"
ORIGIN
        1 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
       61 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
      121 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
      181 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
      241 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
      301 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
      361 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
      421 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
      481 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
      541 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
      601 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
      661 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
      721 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
      781 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      841 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1501 atgctcgtca ggggggcgga gcctgtggaa aaacgccagc aacgcggcct ttttacggtt
     1561 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1621 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1681 gcgcagcgag tcagtgagcg aggaagcctg cataacgcga agtaatcttt tcggttttaa
     1741 agaaaaaggg cagggtggtg acaccttgcc cttttttgcc ggactgcagc ggccgctagc
     1801 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
     1861 tttaagaagg agataaacaa tgggtaaagg agaagaactt ttcactggag ttgtcccaat
     1921 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     1981 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     2041 tgttccgtgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcccgtta
     2101 tccggatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     2161 ggaacgcact atatctttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt
     2221 tgaaggtgat acccttgtta atcgtatcga gttaaaaggt attgatttta aagaagatgg
     2281 aaacattctc ggacacaaac tcgagtacaa ctataactca cacaatgtat acatcacggc
     2341 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg
     2401 atccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     2461 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaatgaaaa
     2521 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     2581 tgaactatac aagtggggtc ttagcgcggg cgccgtagat ttgagaactt tgggatattc
     2641 acagcagcag caggaaaaga tcaagcccaa agtgaggtcg acagtcgcgc agcatcacga
     2701 agcgctggtg ggtcatgggt ttacacatgc ccacatcgta gccttgtcgc agcaccctgc
     2761 agcccttggc acggtcgccg tcaagtacca ggacatgatt gcggcgttgc cggaagccac
     2821 acatgaggcg atcgtcggtg tggggaaaca gtggagcgga gcccgagcgc ttgaggccct
     2881 gttgacggtc gcgggagagc tgagagggcc tccccttcag ctggacacgg gccagttgct
     2941 gaagatcgcg aagcggggag gagtcacggc ggtcgaggcg gtgcacgcgt ggcgcaatgc
     3001 gctcacggga gcacccctca acctgacccc agagcaggtc gtggccattg cctcgaatgg
     3061 aggtggcaaa caggctctgg aaaccgtaca acgattgctg ccggtgctgt gccaagcgca
     3121 cgggttgacc ccagagcagg tcgtggcgat cgcaagccac gacggtggca aacaggctct
     3181 ggaaacagta cagaggctgt tgcctgtgct gtgccaagcg cacggactta ccccagagca
     3241 ggtcgtggca atcgcctcca acattggtgg caaacaggct ctggaaactg tccagcgcct
     3301 gcttcccgtg ctgtgccaag cgcacggttt aaccccagag caggtcgtgg caatcgcgag
     3361 caataacggt ggcaaacagg ctctggaaac ggtgcagagg ctccttccag tgctgtgcca
     3421 agcgcacggg ctcaccccag agcaggtcgt ggcaatcgcc tccaacattg gtggcaaaca
     3481 ggctctggaa actgtccagc gcctgcttcc cgtgctgtgc caagcgcacg gtctgacccc
     3541 agagcaggtc gtggccattg cctcgaatgg aggtggcaaa caggctctgg aaaccgtaca
     3601 acgattgctg ccggtgctgt gccaagcgca cgggttaacc ccagagcagg tcgtggcaat
     3661 cgcgagcaat aacggtggca aacaggctct ggaaacggtg cagaggctcc ttccagtgct
     3721 gtgccaagcg cacggactca ccccagagca ggtcgtggca atcgcctcca acattggtgg
     3781 caaacaggct ctggaaactg tccagcgcct gcttcccgtg ctgtgccaag cgcacggttt
     3841 gaccccagag caggtcgtgg ccattgcctc gaatggaggt ggcaaacagg ctctggaaac
     3901 cgtacaacga ttgctgccgg tgctgtgcca agcgcacggg ttgaccccag agcaggtcgt
     3961 ggcgatcgca agccacgacg gtggcaaaca ggctctggaa acagtacaga ggctgttgcc
     4021 tgtgctgtgc caagcgcacg gacttacccc agagcaggtc gtggcaatcg cgagcaataa
     4081 cggtggcaaa caggctctgg aaacggtgca gaggctcctt ccagtgctgt gccaagcgca
     4141 cggactgacc ccagagcagg tcgtggcgat cgcaagccac gacggtggca aacaggctct
     4201 ggaaacagta cagaggctgt tgcctgtgct gtgccaagcg cacggcctga ccccagagca
     4261 ggtcgtggcc attgcctcga atggaggtgg caaacaggct ctggaaaccg tacaacgatt
     4321 gctgccggtg ctgtgccaag cgcacggttt aaccccagag caggtcgtgg caatcgcgag
     4381 caataacggt ggcaaacagg ctctggaaac ggtgcagagg ctccttccag tgctgtgcca
     4441 agcgcacggg ctcaccccag agcaggtcgt ggccattgcc tcgaatggag gtggcaaaca
     4501 ggctctggaa accgtacaac gattgctgcc ggtgctgtgc caagcgcacg gtctgacccc
     4561 agagcaggtc gtggcgatcg caagccacga cggtggcaaa caggctctgg aaacagtaca
     4621 gaggctgttg cctgtgctgt gccaagcgca cgggttaacc ccagagcagg tcgtggcaat
     4681 cgcgagcaat aacggtggca aacaggctct ggaaacggtg cagaggctcc ttccagtgct
     4741 gtgccaagcg cacggactca ccccagagca ggtcgtggcc attgcctcga atggaggtgg
     4801 caaacaggct ctggaaaccg tacaacgatt gctgccggtg ctgtgccaag cgcacgggtt
     4861 gaccccagag caggtcgtgg ccattgcctc gaatggaggt ggcaaacagg ctctggaaac
     4921 cgtacaacga ttgctgccgg tgctgtgcca agcgcacgga cttaccccag agcaggtcgt
     4981 ggcgatcgca agccacgacg gtggcaaaca ggctctggaa acagtacaga ggctgttgcc
     5041 tgtgctgtgc caagcgcacg gactgagcgc gggcgccgta gatttgagaa ctttgggata
     5101 ttcacagcag cagcaggaaa agatcaagcc caaagtgagg tcgacagtcg cgcagcatca
     5161 cgaagcgctg gtgggtcatg ggtttacaca tgcccacatc gtagccttgt cgcagcaccc
     5221 tgcagccctt ggcacggtcg ccgtcaagta ccaggacatg attgcggcgt tgccggaagc
     5281 cacacatgag gcgatcgtcg gtgtggggaa acagtggagc ggagcccgag cgcttgaggc
     5341 cctgttgacg gtcgcgggag agctgagagg gcctcccctt cagctggaca cgggccagtt
     5401 gctgaagatc gcgaagcggg gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa
     5461 tgcgctcacg ggagcacccc tcaacctgac cccagagcag gtcgtggcaa tcgcctccaa
     5521 cattggtggc aaacaggctc tggaaactgt ccagcgcctg cttcccgtgc tgtgccaagc
     5581 gcacggttta accccagagc aggtcgtggc aatcgcgagc aataacggtg gcaaacaggc
     5641 tctggaaacg gtgcagaggc tccttccagt gctgtgccaa gcgcacgggc tcaccccaga
     5701 gcaggtcgtg gcgatcgcaa gccacgacgg tggcaaacag gctctggaaa cagtacagag
     5761 gctgttgcct gtgctgtgcc aagcgcacgg gttaacccca gagcaggtcg tggccattgc
     5821 ctcgaatgga ggtggcaaac aggctctgga aaccgtacaa cgattgctgc cggtgctgtg
     5881 ccaagcgcac gggttgaccc cagagcaggt cgtggcaatc gcgagcaata acggtggcaa
     5941 acaggctctg gaaacggtgc agaggctcct tccagtgctg tgccaagcgc acggacttac
     6001 cccagagcag gtcgtggcaa tcgcgagcaa taacggtggc aaacaggctc tggaaacggt
     6061 gcagaggctc cttccagtgc tgtgccaagc gcacggactg accccagagc aggtcgtggc
     6121 cattgcctcg aatggaggtg gcaaacaggc tctggaaacc gtacaacgat tgctgccggt
     6181 gctgtgccaa gcgcacggcc tgaccccaga gcaggtcgtg gcaatcgcct ccaacattgg
     6241 tggcaaacag gctctggaaa ctgtccagcg cctgcttccc gtgctgtgcc aagcgcacgg
     6301 tttaacccca gagcaggtcg tggccattgc ctcgaatgga ggtggcaaac aggctctgga
     6361 aaccgtacaa cgattgctgc cggtgctgtg ccaagcgcac gggctcaccc cagagcaggt
     6421 cgtggccatt gccacgaatg gaggtggcaa acaggctctg gaaaccgtac aacgattgct
     6481 gccggtgctg tgccaagcgc acggcctaac cccagagcag gtcgtggcca ttgcctcgaa
     6541 tggaggtggc aaacaggctc tggaaaccgt acaacgattg ctgccggtgc tgtgccaagc
     6601 gcacggtctg accccagagc aggtcgtggc gatcgcaagc cacgacggtg gcaaacaggc
     6661 tctggaaaca gtacagaggc tgttgcctgt gctgtgccaa gcgcacgggt taaccccaga
     6721 gcaggtcgtg gcgatcgcaa gccacgacgg tggcaaacag gctctggaaa cagtacagag
     6781 gctgttgcct gtgctgtgcc aagcgcacgg actcacccca gagcaggtcg tggccattgc
     6841 ctcgaatgga ggtggcaaac aggctctgga aaccgtacaa cgattgctgc cggtgctgtg
     6901 ccaagcgcac gggttgaccc cagagcaggt cgtggcgatc gcaagccacg acggtggcaa
     6961 acaggctctg gaaacagtac agaggctgtt gcctgtgctg tgccaagcgc acggacttac
     7021 cccagagcag gtcgtggcga tcgcaagcca cgacggtggc aaacaggctc tggaaacagt
     7081 acagaggctg ttgcctgtgc tgtgccaagc gcacggactg accccagagc aggtcgtggc
     7141 aatcgcctcc aacattggtg gcaaacaggc tctggaaact gtccagcgcc tgcttcccgt
     7201 gctgtgccaa gcgcacggcc tgaccccaga gcaggtcgtg gcaatcgcct ccaacattgg
     7261 tggcaaacag gctctggaaa ctgtccagcg cctgcttccc gtgctgtgcc aagcgcacgg
     7321 tttaacccca gagcaggtcg tggccattgc ctcgaatgga ggtggcaaac aggctctgga
     7381 aaccgtacaa cgattgctgc cggtgctgtg ccaagcgcac gggctcaccc cagagcaggt
     7441 cgtggcaatc gcctccaaca ttggtggcaa acaggctctg gaatctatcg ttgctcagct
     7501 gtcacgtccg gacccggctc tggctgctct gaccaacgac cacctggttg ctctggcttg
     7561 cctgggtggt cgtccggcta tggacgctgt taaaaaaggc ctagtgccgc gcggcagcca
     7621 ccaccatcac catcattaat tccagaaatc atccttagcg aaagctaagg atttttttta
     7681 tctgaaattc tgcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7741 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7861 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7921 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga a
//
