LOCUS       sequence-305689-        7247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Halothiobacillus neapolitanus CsoS2 with N-terminal His-tag, pET-14b
            backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7247)
  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 7247)
  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: pBz118_His_CsoS2
FEATURES             Location/Qualifiers
     source          1..7247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     127..146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        127..145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             177..199
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             212..229
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2903..2921)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2917..2964
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3330..3335,3336..3352,3353..3358))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3385..3403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3471..3575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3576..3644,3645..4436)
                     /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(3794..3813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4607..5195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5096..5115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5349..5366
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5381..5520
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5446..5465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5606..5628
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5622..5813)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
       61 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
      121 cgaaattaat acgactcact atagggacac cacaacggtt tccctctaga aataattttg
      181 tttaacttta agaaggagat ataccatgtc ccaccatcat catcatcatc cttcacagtc
      241 aggaatgaat cctgccgacc tgagcggact ctctggcaag gaactggcac gcgcacgccg
      301 cgctgcacta tccaagcaag ggaaagcagc tgtttctaat aaaacggcta gcgtaaaccg
      361 tagcactaaa caggcggcat cttcgatcaa tacaaatcag gtgcgctctt ctgtaaatga
      421 agtgcccact gattaccaaa tggcggatca attgtgctct acgattgatc atgctgactt
      481 tggtaccgaa agcaatcgcg ttagagatct ctgccgtcaa cgcagagagg cactatcaac
      541 tatcggtaaa aaagcggtta aaaccaacgg caagccgtcg ggtcgcgttc gaccacagca
      601 atcagtggtt cacaacgacg caatgatcga aaatgccggt gatactaacc aatcatcgtc
      661 cacttcatta aataatgaac tttccgaaat ctgctccata gcagacgaca tgccggagcg
      721 ttttggttca caagccaaaa ccgtccgtga tatctgccgt gcacgccgtc aagcgctctc
      781 tgagcgtgga actcgcgccg tgccgccaaa gccgcaatct caaggtggtc caggacgcaa
      841 tggctatcaa attgatggat acctagatac cgcacttcat ggccgcgatg ccgccaagcg
      901 ccaccgtgaa atgctctgtc aatacggccg cggcacagca ccttcctgca agccaacagg
      961 ccgtgtcaaa aattctgtac agtcgggcaa cgcagcgcca aaaaaggttg aaaccggtca
     1021 caccctatcc ggcggatctg ttacgggcac gcaagtggat cgtaaatctc atgtgactgg
     1081 aaacgagccg ggcacttgcc gagcagtcac gggcaccgag tacgtaggta ctgagcaatt
     1141 cacctctttt tgcaatacca gccccaagcc aaatgcgacg aaggtcaatg tgaccacaac
     1201 ggctcgtggt cgccctgtta gcggtacgga agtttcacgg accgagaagg taactggcaa
     1261 cgaatccggt gtctgccgta acgttaccgg caccgaatac atgagtaatg aagctcactt
     1321 ttctctatgt ggaacagccg caaagccttc acaagcggat aaagtcatgt tcggcgccac
     1381 agcacgaacg catcaagtgg tcagtggcag tgatgaattc aggccctctt ctgttacggg
     1441 taacgaatcg ggtgcaaaac gcacaattac cggctcgcag tacgcagacg aaggtcttgc
     1501 gcgactcacg atcaacggag cacctgcaaa agtagccaga acccacacct ttgcgggctc
     1561 tgacgttacc ggcacggaaa tcggtcgctc tactcgcgta actggtgatg aaagcggttc
     1621 gtgtcgttca atctcaggca ccgagtatct cagtaacgag caattccaat ccttttgtga
     1681 cacaaaacct caacgcagcc cgttcaaggt tggccaagat cgcacgaaca agggtcagtc
     1741 tgtgactggt aacttggttg atcgttccga actggttaca ggtaacgaac caggttcatg
     1801 ctctcgggtt acaggctctc agtatggcca aagcaaaatc tgcggtggtg gcgtgggaaa
     1861 agtgcgctca atgcgcaccc ttcgcggcac ctcagtatct ggccaacagc tagatcatgc
     1921 cccaaagatg tccggtgacg agcgcggcgg gtgcatgccc gtcaccggta atgagtacta
     1981 cggtcgtgaa catttcgaac cgttttgtac gagcacccca gagcccgaag ctcaatcaac
     2041 tgaacaatca ttgacctgtg aaggacaaat tattagcggc acttcagttg acgccagtga
     2101 tttggtcaca ggaaatgaaa tcggtgaaca gcaactcatc agcggtgacg cctatgttgg
     2161 cgcgcagcag acaggttgcc ttcccactag tccacgcttc aaccaaactg gcaatgttca
     2221 gtcaatgggt tttaagaaca ccaatcagcc agaacaaaac tttgcaccag gtgaagtaat
     2281 gcctactgac tttagtattc aaaccccagc tcgctcggct cagaatcgca ttacaggtaa
     2341 cgacattgcg ccctcaggtc gcattacagg ccctggtatg ctggcaaccg gcttgattac
     2401 aggaaccccc gaattcaggc acgctgcgcg cgagttggtt ggttctccac aacccatggc
     2461 aatggccatg gccaaccgta ataaggctgc tcaagcacct gttgtgcagc cagaagtggt
     2521 tgcaactcag gaaaagcctg agttggtatg tgcaccaaga agcgatcaaa tggatcgtgt
     2581 gagtggcgaa ggcaaagaac gttgccacat cactggcgat gactggtcag ttaacaagca
     2641 catcaccggt acagccggtc aatgggcgag tggtcgcaac ccttccatgc gcggtaatgc
     2701 gcgtgtggtc gaaaccagcg cgtttgccaa tcgcaatgtg ccaaaacctg aaaagccggg
     2761 ctccaagatc acgggcagta gtggtaatga cacccaaggt agtctgatca cttactccgg
     2821 cggcgcgcgc ggttgattaa gtaaagcggt taagatccgg ctgctaacaa agcccgaaag
     2881 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2941 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac
     3001 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
     3061 tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat
     3121 caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat
     3181 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
     3241 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
     3301 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
     3361 atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3421 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     3841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4081 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     4141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4261 ttgctgataa atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc
     4321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5401 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     5461 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     5521 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5581 gcttacagac aagctgtgac cgcctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5641 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     5701 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5761 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     5821 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     5881 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     5941 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     6001 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     6061 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     6121 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     6181 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     6241 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     6301 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     6361 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     6421 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     6481 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     6541 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg
     6601 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     6661 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     6721 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     6781 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     6841 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     6901 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     6961 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     7021 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct
     7081 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     7141 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     7201 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaag
//
