LOCUS       sequence-300557-        5676 bp    DNA     circular SYN 15-APR-2025
DEFINITION  His-tagged CsoS2 Middle Region (MR), pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  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 5676)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pBz106_CsoS2_MR
FEATURES             Location/Qualifiers
     source          1..5676
                     /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             67..84
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1187..1205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1201..1248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1614..1619,1620..1636,1637..1642))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1669..1687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1755..1859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1860..1928,1929..2720)
                     /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(2078..2097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2891..3479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3380..3399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3633..3650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3665..3804
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3730..3749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3890..3912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3906..4097)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5555..5574
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5658..5676,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5658..5676
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
       61 atgtcccacc atcatcatca tcatgcgcca aaaaaggttg aaaccggtca caccctatcc
      121 ggcggatctg ttacgggcac gcaagtggat cgtaaatctc atgtgactgg aaacgagccg
      181 ggcacttgcc gagcagtcac gggcaccgag tacgtaggta ctgagcaatt cacctctttt
      241 tgcaatacca gccccaagcc aaatgcgacg aaggtcaatg tgaccacaac ggctcgtggt
      301 cgccctgtta gcggtacgga agtttcacgg accgagaagg taactggcaa cgaatccggt
      361 gtctgccgta acgttaccgg caccgaatac atgagtaatg aagctcactt ttctctatgt
      421 ggaacagccg caaagccttc acaagcggat aaagtcatgt tcggcgccac agcacgaacg
      481 catcaagtgg tcagtggcag tgatgaattc aggccctctt ctgttacggg taacgaatcg
      541 ggtgcaaaac gcacaattac cggctcgcag tacgcagacg aaggtcttgc gcgactcacg
      601 atcaacggag cacctgcaaa agtagccaga acccacacct ttgcgggctc tgacgttacc
      661 ggcacggaaa tcggtcgctc tactcgcgta actggtgatg aaagcggttc gtgtcgttca
      721 atctcaggca ccgagtatct cagtaacgag caattccaat ccttttgtga cacaaaacct
      781 caacgcagcc cgttcaaggt tggccaagat cgcacgaaca agggtcagtc tgtgactggt
      841 aacttggttg atcgttccga actggttaca ggtaacgaac caggttcatg ctctcgggtt
      901 acaggctctc agtatggcca aagcaaaatc tgcggtggtg gcgtgggaaa agtgcgctca
      961 atgcgcaccc ttcgcggcac ctcagtatct ggccaacagc tagatcatgc cccaaagatg
     1021 tccggtgacg agcgcggcgg gtgcatgccc gtcaccggta atgagtacta cggtcgtgaa
     1081 catttcgaac cgttttgtac gagcacccca gagcccgaag ctcaataaag cggttaagat
     1141 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1201 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1261 actatatccg gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc
     1321 tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag
     1381 aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact
     1441 ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa
     1501 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     1561 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     1621 cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac
     1681 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1741 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1801 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1861 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1921 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1981 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2041 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2101 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2161 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2221 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2281 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2341 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2401 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2461 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2521 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggctctc
     2581 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2641 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2701 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2761 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2821 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3181 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3241 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3301 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3361 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3421 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3481 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3541 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3601 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3661 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3721 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3781 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3841 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgcctc cgggagctgc
     3901 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3961 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4021 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4081 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4141 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4201 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4261 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4321 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4381 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4441 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4501 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4561 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     4621 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     4681 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     4741 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     4801 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     4861 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     4921 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     4981 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     5041 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     5101 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     5161 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     5221 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5281 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5341 ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5401 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5461 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5521 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     5581 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     5641 tctcgatccc gcgaaattaa tacgactcac tatagg
//
