LOCUS       sequence-297981-        5781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged first two N-peptides of NTD fused to Superfolder GFP 
            ([N1-N2]-GFP) used for MST, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  AUTHORS   Oltrogge LM, Chaijarasphong T, Chen AW, Bolin ER, Marqusee S, Savage
            DF
  TITLE     Multivalent interactions between CsoS2 and Rubisco mediate 
            alpha-carboxysome formation.
  JOURNAL   Nat Struct Mol Biol. 2020 Mar;27(3):281-287. doi: 
            10.1038/s41594-020-0387-7. Epub 2020 Mar 2.
  PUBMED    32123388
REFERENCE   2  (bases 1 to 5781)
  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"; doi: 
            "10.1038/s41594-020-0387-7"; journalName: "Nat Struct Mol Biol"; 
            date: "2020-03"; volume: "27"; issue: "3"; pages: "281-287"
COMMENT     Sequence Label: pLz26_CsoS2_N1-N2_GFP
FEATURES             Location/Qualifiers
     source          1..5781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             19..39
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1144..1167
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1229..1247)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1243..1290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1656..1661,1662..1678,1679..1684))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1711..1729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1797..1901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1902..1970,1971..2762)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2120..2139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2933..3521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3422..3441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3675..3692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3707..3846
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3772..3791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3932..3954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3948..4139)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5597..5616
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5700..5719
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5700..5718
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5750..5772
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 caccatcacc atcaccatga aaacctgtac ttccagtccc cttcacagtc aggaatgaat
       61 cctgccgacc tgagcggact ctctggcaag gaactggcac gcgcacgccg cgctgcacta
      121 tccaagcaag ggaaagcagc tgtttctaat aaaacggcta gcgtaaaccg tagcactaaa
      181 caggcggcat cttcgatcaa tacaaatcag gtgcgctctt ctgtaaatga agtgcccact
      241 gattaccaaa tggcggatca attgtgctct acgattgatc atgctgactt tggtaccgaa
      301 agcaatcgcg ttagagatct ctgccgtcaa cgcagagagg cactatcaac tatcggtaaa
      361 aaagcggtta aaaccaacgg caagccgtcg ggtcgcgttc gaccacagca atcaggtagc
      421 ggaagcggcg gttctaaagg tgaagaactg ttcaccggtg ttgttccgat cctggttgaa
      481 ctggatggtg atgttaacgg ccacaaattc tctgttcgtg gtgaaggtga aggtgatgca
      541 accaacggta aactgaccct gaaattcatc tgcactaccg gtaaactgcc ggttccatgg
      601 ccgactctgg tgactaccct gacctatggt gttcagtgtt tttctcgtta cccggatcac
      661 atgaagcagc atgatttctt caaatctgca atgccggaag gttatgtaca ggagcgcacc
      721 atttctttca aagacgatgg cacctacaaa acccgtgcag aggttaaatt tgaaggtgat
      781 actctggtga accgtattga actgaaaggc attgatttca aagaggacgg caacatcctg
      841 ggccacaaac tggaatataa cttcaactcc cataacgttt acatcaccgc agacaaacag
      901 aagaacggta tcaaagctaa cttcaaaatt cgccataacg ttgaagacgg tagcgtacag
      961 ctggcggacc actaccagca gaacactccg atcggtgatg gtccggttct gctgccggat
     1021 aaccactacc tgtccaccca gtctaaactg tccaaagacc cgaacgaaaa gcgcgaccac
     1081 atggtgctgc tggagttcgt tactgcagca ggtatcacgc acggcatgga tgagctctac
     1141 aaatggagtc atccacagtt tgaaaagtaa agcggttaag atccggctgc taacaaagcc
     1201 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1261 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatcca
     1321 caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag
     1381 caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa
     1441 ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg
     1501 gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc
     1561 cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg
     1621 actgcgttag caatttaact gtgataaact accgcattaa agcttatcga tgataagctg
     1681 tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1741 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1801 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1861 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1921 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1981 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2041 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2101 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2161 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2221 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2281 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2341 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2401 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2461 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2521 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2581 ggtttattgc tgataaatct ggagccggtg agcgtggctc tcgcggtatc attgcagcac
     2641 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2701 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2761 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3061 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3601 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3661 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3721 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3781 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3841 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3901 gcatccgctt acagacaagc tgtgaccgcc tccgggagct gcatgtgtca gaggttttca
     3961 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4021 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4081 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4141 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4201 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     4261 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     4321 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     4381 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4441 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4501 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4561 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     4621 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     4681 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     4741 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     4801 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     4861 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     4921 tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt
     4981 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     5041 gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc
     5101 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     5161 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     5221 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     5281 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     5341 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     5401 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     5461 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     5521 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     5581 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     5641 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     5701 aatacgactc actataggga caccacaacg gtttccctct agaaataatt ttgtttaact
     5761 ttaagaagga gatataccat g
//
