LOCUS       sequence-298011-        5622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged fourth N-peptide of NTD fused to Superfolder GFP 
            ([N4]-GFP) used for BLI, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  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 5622)
  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: pBz175_CsoS2_N4_GFP
FEATURES             Location/Qualifiers
     source          1..5622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             86..103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             314..334
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1067..1090
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1152..1170)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1166..1213
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1579..1584,1585..1601,1602..1607))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1634..1652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1720..1824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1825..1893,1894..2685)
                     /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(2043..2062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2856..3444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3345..3364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3598..3615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3630..3769
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3695..3714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3855..3877
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3871..4062)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5520..5539
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg acaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgtcccacca tcatcatcat catagagcag ttccaccaaa
      121 gccgcaatct caaggtggtc caggacgcaa tggctatcaa attgatggat acctagatac
      181 cgcacttcat ggccgcgatg ccgccaagcg ccaccgtgaa atgctctgtc aatacggccg
      241 cggcacagca ccttcctgca agccaacagg ccgtgtcaaa aattctgtac agtcgggcaa
      301 cgcagcgcca aaagaaaacc tgtacttcca gtcctcaggt agcggaagcg gcggttctaa
      361 aggtgaagaa ctgttcaccg gtgttgttcc gatcctggtt gaactggatg gtgatgttaa
      421 cggccacaaa ttctctgttc gtggtgaagg tgaaggtgat gcaaccaacg gtaaactgac
      481 cctgaaattc atctgcacta ccggtaaact gccggttcca tggccgactc tggtgactac
      541 cctgacctat ggtgttcagt gtttttctcg ttacccggat cacatgaagc agcatgattt
      601 cttcaaatct gcaatgccgg aaggttatgt acaggagcgc accatttctt tcaaagacga
      661 tggcacctac aaaacccgtg cagaggttaa atttgaaggt gatactctgg tgaaccgtat
      721 tgaactgaaa ggcattgatt tcaaagagga cggcaacatc ctgggccaca aactggaata
      781 taacttcaac tcccataacg tttacatcac cgcagacaaa cagaagaacg gtatcaaagc
      841 taacttcaaa attcgccata acgttgaaga cggtagcgta cagctggcgg accactacca
      901 gcagaacact ccgatcggtg atggtccggt tctgctgccg gataaccact acctgtccac
      961 ccagtctaaa ctgtccaaag acccgaacga aaagcgcgac cacatggtgc tgctggagtt
     1021 cgttactgca gcaggtatca cgcacggcat ggatgagctc tacaaatgga gtcatccaca
     1081 gtttgaaaag taaagcggtt aagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1141 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1201 gaggggtttt ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg
     1261 ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc
     1321 caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata
     1381 gcgctagcag cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga
     1441 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga
     1501 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta
     1561 actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct
     1621 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1681 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1741 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1801 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1861 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1921 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1981 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2041 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     2101 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2161 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2221 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2281 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2341 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     2401 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2461 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2521 tctggagccg gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag
     2581 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2641 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2701 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2761 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2821 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2881 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2941 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3001 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3061 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3121 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3181 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3241 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3301 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3361 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3421 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3481 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3541 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3601 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     3661 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3721 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3781 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3841 agctgtgacc gcctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3901 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3961 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4021 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     4081 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     4141 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     4201 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     4261 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     4321 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     4381 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     4441 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     4501 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg
     4561 cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc
     4621 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
     4681 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     4741 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
     4801 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
     4861 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
     4921 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
     4981 aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc
     5041 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     5101 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     5161 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     5221 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     5281 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc
     5341 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5401 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5461 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5521 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     5581 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
