LOCUS       sequence-298012-        6165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged CsoS2 N-terminal domain (NTD) fused to Superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  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 6165)
  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: pLz33_CsoS2_NTD_GFP
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             32..54
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             64..81
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             82..102
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1676..1694)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1690..1737
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2103..2108,2109..2125,2126..2131))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2158..2176)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2244..2348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2349..2417,2418..3209)
                     /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(2567..2586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3380..3968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3869..3888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4122..4139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4154..4293
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4219..4238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4379..4401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4395..4586)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6044..6063
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6147..6165,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6147..6165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
       61 atgcaccatc accatcacca tgaaaacctg tacttccagt ccccttcaca gtcaggaatg
      121 aatcctgccg acctgagcgg actctctggc aaggaactgg cacgcgcacg ccgcgctgca
      181 ctatccaagc aagggaaagc agctgtttct aataaaacgg ctagcgtaaa ccgtagcact
      241 aaacaggcgg catcttcgat caatacaaat caggtgcgct cttctgtaaa tgaagtgccc
      301 actgattacc aaatggcgga tcaattgtgc tctacgattg atcatgctga ctttggtacc
      361 gaaagcaatc gcgttagaga tctctgccgt caacgcagag aggcactatc aactatcggt
      421 aaaaaagcgg ttaaaaccaa cggcaagccg tcgggtcgcg ttcgaccaca gcaatcagtg
      481 gttcacaacg acgcaatgat cgaaaatgcc ggtgatacta accaatcatc gtccacttca
      541 ttaaataatg aactttccga aatctgctcc atagcagacg acatgccgga gcgttttggt
      601 tcacaagcca aaaccgtccg tgatatctgc cgtgcacgcc gtcaagcgct ctctgagcgt
      661 ggaactcgcg ccgtgccgcc aaagccgcaa tctcaaggtg gtccaggacg caatggctat
      721 caaattgatg gatacctaga taccgcactt catggccgcg atgccgccaa gcgccaccgt
      781 gaaatgctct gtcaatacgg ccgcggcaca gcaccttcct gcaagccaac aggccgtgtc
      841 aaaaattctg tacagtcggg caacgcagcg ccaaaaaagg tttcaggtag cggaagcggc
      901 ggttctaaag gtgaagaact gttcaccggt gttgttccga tcctggttga actggatggt
      961 gatgttaacg gccacaaatt ctctgttcgt ggtgaaggtg aaggtgatgc aaccaacggt
     1021 aaactgaccc tgaaattcat ctgcactacc ggtaaactgc cggttccatg gccgactctg
     1081 gtgactaccc tgacctatgg tgttcagtgt ttttctcgtt acccggatca catgaagcag
     1141 catgatttct tcaaatctgc aatgccggaa ggttatgtac aggagcgcac catttctttc
     1201 aaagacgatg gcacctacaa aacccgtgca gaggttaaat ttgaaggtga tactctggtg
     1261 aaccgtattg aactgaaagg cattgatttc aaagaggacg gcaacatcct gggccacaaa
     1321 ctggaatata acttcaactc ccataacgtt tacatcaccg cagacaaaca gaagaacggt
     1381 atcaaagcta acttcaaaat tcgccataac gttgaagacg gtagcgtaca gctggcggac
     1441 cactaccagc agaacactcc gatcggtgat ggtccggttc tgctgccgga taaccactac
     1501 ctgtccaccc agtctaaact gtccaaagac ccgaacgaaa agcgcgacca catggtgctg
     1561 ctggagttcg ttactgcagc aggtatcacg cacggcatgg atgagctcta caaataaagc
     1621 ggttaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1681 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1741 aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc
     1801 gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag
     1861 tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc
     1921 atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg
     1981 cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc
     2041 attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     2101 gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc
     2161 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2221 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2281 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2341 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2401 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2461 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2521 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2581 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2641 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2701 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2761 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2821 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2881 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2941 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3001 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3061 gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3121 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3181 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3241 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3301 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3361 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3421 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3481 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3541 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3601 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3661 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3721 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3781 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3841 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3901 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3961 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4021 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4081 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4141 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4201 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     4261 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4321 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgcctcc
     4381 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     4441 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     4501 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     4561 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     4621 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4681 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4741 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     4801 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4861 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4921 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4981 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5041 atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag
     5101 atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc
     5161 aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt
     5221 ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa
     5281 ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat
     5341 aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg
     5401 atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg
     5461 cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc
     5521 gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg
     5581 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     5641 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     5701 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     5761 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     5821 ggttgaaggc tctcaagggc atcggtcgac gctctccctt atgcgactcc tgcattagga
     5881 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     5941 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6001 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6061 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6121 ggatcgagat ctcgatcccg cgaaattaat acgactcact atagg
//
