LOCUS       sequence-300476-        5685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged first N-peptide of NTD fused to Superfolder GFP 
            ([N1]-GFP) used for BLI and MST, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5685)
  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 5685)
  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: pBz172_CsoS2_N1_GFP
FEATURES             Location/Qualifiers
     source          1..5685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             304..324
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1057..1080
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1142..1160)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1156..1203
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1569..1574,1575..1591,1592..1597))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1624..1642)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1710..1814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1815..1883,1884..2675)
                     /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(2033..2052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2846..3434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3335..3354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3588..3605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3620..3759
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3685..3704)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3845..3867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3861..4052)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5510..5529
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5613..5632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5613..5631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5663..5685
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tataccatgt cccaccatca tcatcatcat ccttcacagt caggaatgaa tcctgccgac
       61 ctgagcggac tctctggcaa ggaactggca cgcgcacgcc gcgctgcact atccaagcaa
      121 gggaaagcag ctgtttctaa taaaacggct agcgtaaacc gtagcactaa acaggcggca
      181 tcttcgatca atacaaatca ggtgcgctct tctgtaaatg aagtgcccac tgattaccaa
      241 atggcggatc aattgtgctc tacgattgat catgctgact ttggtaccga aagcaatcgc
      301 gttgaaaacc tgtacttcca gtcctcaggt agcggaagcg gcggttctaa aggtgaagaa
      361 ctgttcaccg gtgttgttcc gatcctggtt gaactggatg gtgatgttaa cggccacaaa
      421 ttctctgttc gtggtgaagg tgaaggtgat gcaaccaacg gtaaactgac cctgaaattc
      481 atctgcacta ccggtaaact gccggttcca tggccgactc tggtgactac cctgacctat
      541 ggtgttcagt gtttttctcg ttacccggat cacatgaagc agcatgattt cttcaaatct
      601 gcaatgccgg aaggttatgt acaggagcgc accatttctt tcaaagacga cggcacctac
      661 aaaacccgtg cagaggttaa atttgaaggt gatactctgg tgaaccgtat tgaactgaaa
      721 ggcattgatt tcaaagagga cggcaacatc ctgggccaca aactggaata taacttcaac
      781 tcccataacg tttacatcac cgcagacaaa cagaagaacg gtatcaaagc taacttcaaa
      841 attcgccata acgttgaaga cggtagcgta cagctggcgg accactacca gcagaacact
      901 ccgatcggtg atggtccggt tctgctgccg gataaccact acctgtccac ccagtctaaa
      961 ctgtccaaag acccgaacga aaagcgcgac cacatggtgc tgctggagtt cgttactgca
     1021 gcaggtatca cgcacggcat ggatgagctc tacaaatgga gtcatccaca gtttgaaaag
     1081 taaagcggtt aagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1141 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1201 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
     1261 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     1321 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     1381 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     1441 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     1501 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     1561 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     1621 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1681 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1741 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1801 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1861 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1921 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1981 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2041 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2101 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2161 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2221 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2281 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2341 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2401 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2461 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2521 gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2581 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2641 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2701 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2761 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2821 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2881 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2941 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3001 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3061 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3121 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3181 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3241 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3301 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3361 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3421 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3481 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3541 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3601 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3661 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3721 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3781 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3841 gcctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3901 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3961 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4021 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4081 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4141 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4201 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4261 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4321 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4381 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4441 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4501 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     4561 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     4621 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4681 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4741 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4801 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4861 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4921 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4981 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     5041 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     5101 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     5161 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     5221 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     5281 tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca
     5341 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     5401 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5461 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5521 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5581 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag ggacaccaca
     5641 acggtttccc tctagaaata attttgttta actttaagaa ggaga
//
