LOCUS       sequence-273735-        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     1572..1591
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1675..1694
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1675..1693
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1725..1747
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1766..1783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1784..1804
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2150..2168)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2164..2211
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2577..2582,2583..2599,2600..2605))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2632..2650)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2718..2822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2823..2891,2892..3683)
                     /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(3041..3060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3854..4442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4343..4362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4596..4613
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4628..4767
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4693..4712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4853..4875
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(4869..4946,1..114))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
       61 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
      121 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
      181 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
      241 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
      301 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
      361 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
      421 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
      481 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
      541 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
      601 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
      661 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
      721 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
      781 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
      841 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccagtgatc
      901 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
      961 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     1021 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     1081 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     1141 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     1201 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     1261 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     1321 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgacgc
     1381 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     1441 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     1501 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     1561 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     1621 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     1681 gactcactat agggacacca caacggtttc cctctagaaa taattttgtt taactttaag
     1741 aaggagatat accatgtcct ggaagcacca tcaccatcac catgaaaacc tgtacttcca
     1801 gtccgcagct gttggtgggg gatctggagg tgggtctggt ggacctcccc cgcctccacc
     1861 gccaccgcca gctcctgcac ctgcgcctgc tcctccgcca ccccctccac cgccacctgc
     1921 tcctgctcct gctccggcac caccacctcc gcctccgcca cctggaggtg gatcaggagg
     1981 tggttcaggt ggaggacgtg atttagctcg cgctcgtcgt gaggcattaa gtcaacaggg
     2041 taaagcagcc gtcgggggcg gtagcggagg tggtagtggt ggaagcggtt aagcggttaa
     2101 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2161 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2221 ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt
     2281 ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc
     2341 gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg
     2401 actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac
     2461 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     2521 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     2581 aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga
     2641 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2821 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2881 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2941 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3001 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3061 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3121 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3181 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3241 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3301 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3361 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3421 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3481 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggct
     3541 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3601 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3661 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3721 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3781 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3841 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3901 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3961 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4021 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4081 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4141 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4201 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4261 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4321 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4381 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4441 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4501 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4561 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4621 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     4681 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     4741 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4801 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgc ctccgggagc
     4861 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     4921 tcatcagcgt ggtcgtgaag cgattc
//
