LOCUS       sequence-297997-        4946 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Consensus N*-peptide after polyproline II helix, N-terminal His- tag
            used as BLI bait, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  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 4946)
  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: pLz47_polyPro_CsoS2_Npeptide
FEATURES             Location/Qualifiers
     source          1..4946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..97
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        78..96
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             128..150
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             169..186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             187..207
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(553..571)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      567..614
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(980..985,986..1002,1003..1008))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1035..1053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1121..1225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1226..1294,1295..2086)
                     /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(1444..1463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2257..2845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2746..2765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2999..3016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3031..3170
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3096..3115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3256..3278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3272..3463)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4921..4940
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
       61 tctcgatccc gcgaaattaa tacgactcac tatagggaca ccacaacggt ttccctctag
      121 aaataatttt gtttaacttt aagaaggaga tataccatgt cctggaagca ccatcaccat
      181 caccatgaaa acctgtactt ccagtccgca gctgttggtg ggggatctgg aggtgggtct
      241 ggtggacctc ccccgcctcc accgccaccg ccagctcctg cacctgcgcc tgctcctccg
      301 ccaccccctc caccgccacc tgctcctgct cctgctccgg caccaccacc tccgcctccg
      361 ccacctggag gtggatcagg aggtggttca ggtggaggac gtgatttagc tcgcgctcgt
      421 cgtgaggcat taagtcaaca gggtaaagca gccgtcgggg gcggtagcgg aggtggtagt
      481 ggtggaagcg gttaagcggt taagatccgg ctgctaacaa agcccgaaag gaagctgagt
      541 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      601 tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac gggtgtggtc
      661 gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac tgggcggcgg
      721 ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat caacgcatat
      781 agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat atcccgcaag
      841 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg
      901 aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt
      961 aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc
     1021 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1081 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1141 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1201 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1261 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1321 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1381 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1441 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1501 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1561 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1621 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1681 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1741 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1801 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1861 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1921 atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc cagatggtaa
     1981 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2041 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2101 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2161 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2341 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2401 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2461 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2521 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2581 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2641 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2701 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2761 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2821 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2881 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2941 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3001 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3061 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3121 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3181 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3241 aagctgtgac cgcctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3301 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3361 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3421 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3481 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     3541 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     3601 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     3661 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     3721 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     3781 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     3841 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     3901 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc
     3961 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
     4021 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
     4081 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
     4141 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     4201 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     4261 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     4321 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     4381 gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc
     4441 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     4501 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     4561 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4621 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4681 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct ctcccttatg
     4741 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     4801 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     4861 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     4921 ggtgatgtcg gcgatatagg cgccag
//
