LOCUS       sequence-300546-        6426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wild-type Halothiobacillus neapolitanus Form I Rubisco with StrepII 
            tag (CbbL-StrepII, CbbS), pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6426)
  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 6426)
  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: pBz15_Strep_Rubisco
FEATURES             Location/Qualifiers
     source          1..6426
                     /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             1502..1525
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1956..1974)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1970..2017
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2383..2388,2389..2405,2406..2411))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2438..2456)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2524..2628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2629..2697,2698..3489)
                     /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(2847..2866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3660..4248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4149..4168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4402..4419
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4434..4573
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4499..4518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4659..4681
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4675..4866)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6324..6343
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg acaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgtccgcagt taaaaagtat agtgctggtg taaaagaata
      121 ccggcagacc tattggatgc cggaatacac accgttggat tccgacatcc ttgcatgctt
      181 caaaatcacc ccacaaccgg gtgttgatcg cgaagaagcc gcagccgcgg ttgcagcaga
      241 atcttcaacc ggcacatgga ccaccgtgtg gaccgatttg ctgaccgaca tggactacta
      301 caaaggccgt gcctatcgca ttgaagacgt acccggtgac gatgcggcat tctatgcctt
      361 tatcgcctac ccaatcgacc tgttcgaaga agggtcagtt gttaacgtgt ttacctcact
      421 ggttggtaac gtgttcggct tcaaagcggt acgcggcctg cgtctggaag atgttcgctt
      481 cccactcgcc tacgttaaaa cctgtggcgg cccaccgcac ggtattcaag tcgaacgcga
      541 caagatgaac aaatatggtc gcccactgtt gggttgcacc atcaagccaa aacttggttt
      601 gtctgcgaaa aactacggcc gtgccgtata cgagtgcctc cgtggcggcc tcgacttcac
      661 taaagatgat gaaaacatca attctcagcc gttcatgcgc tggcgcgatc gcttcttgtt
      721 cgtacaagac gcgaccgaaa ctgctgaagc ccaaaccggc gaacgcaaag gccattacct
      781 caacgtaacg gcgccaactc ctgaagaaat gtacaagcgc gcagaattcg ccaaagaaat
      841 tggcgcgcca atcattatgc acgactacat caccggtggc ttcacggcca acactggctt
      901 ggccaagtgg tgtcaagaca acggcgtact gctgcacatc caccgtgcga tgcatgcggt
      961 tatcgaccgt aacccgaacc acggtattca cttccgtgtt ctgaccaaga ttctgcgttt
     1021 gtcgggtggc gatcacctgc acaccggtac cgttgtcggc aaactggaag gcgaccgtgc
     1081 ctctactctg ggctggattg atttgctccg cgaatcgttt atccctgaag atcgctcgcg
     1141 cggtatcttc ttcgatcaag actggggttc aatgccaggc gtattcgctg tggcctctgg
     1201 tggtattcac gtatggcaca tgcctgcgct ggttaacatc tttggtgacg actctgtcct
     1261 ccaattcggt ggcggtacgc tgggtcatcc atggggcaac gctgccggtg ctgctgccaa
     1321 ccgtgttgct ctggaagcct gcgtagaagc gcgtaaccaa ggccgcgata tcgaaaaaga
     1381 aggcaaagaa attctgactg ctgctgcaca gcacagccca gaactgaaga ttgccatgga
     1441 aacttggaaa gagatcaaat tcgaatttga cactgtcgac aaactcgaca ctcaaaatcg
     1501 ttggagccac ccgcagttcg aaaagtgatc cctcgtacca cacaacatac taaggtgagt
     1561 aaccatggct gaaatgcagg attacaagca aagcctcaaa tatgagactt tctcttatct
     1621 tccacccatg aacgcggaac gcatccgcgc tcaaatcaag tacgcaattg ctcaaggctg
     1681 gagccccggc attgagcacg tagaagtgaa aaactccatg aaccaatatt ggtacatgtg
     1741 gaaacttccc ttcttcggcg aacaaaatgt cgacaacgtg ttggctgaaa ttgaagcgtg
     1801 tcgtagtgcg tatccaacac accaggtcaa actggtggct tatgacaact atgcgcaaag
     1861 cttaggtctg gccttcgtgg tctaccgcgg caactaaagc ggttaagatc cggctgctaa
     1921 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1981 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2041 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     2101 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     2161 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     2221 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     2281 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     2341 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     2401 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2461 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2521 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2581 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2641 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2701 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2761 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2821 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2881 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2941 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3001 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3061 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3121 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3181 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3241 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3301 gctggctggt ttattgctga taaatctgga gccggtgagc gtggctctcg cggtatcatt
     3361 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3421 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3481 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3541 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3601 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3661 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3721 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3781 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3841 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3901 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3961 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4021 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4081 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4141 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4201 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4261 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4321 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4381 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4441 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4501 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4561 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4621 gctcccggca tccgcttaca gacaagctgt gaccgcctcc gggagctgca tgtgtcagag
     4681 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4741 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4801 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4861 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     4921 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4981 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5041 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5101 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5161 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5221 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5281 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     5341 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     5401 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     5461 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     5521 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     5581 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     5641 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     5701 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     5761 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     5821 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     5881 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     5941 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     6001 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     6061 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6121 atcggtcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     6181 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6241 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6301 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6361 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     6421 cgaaat
//
