LOCUS       sequence-273762-        5781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged first two N-peptides of NTD fused to Superfolder GFP 
            ([N1-N2]-GFP) used for MST, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  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 5781)
  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: pLz26_CsoS2_N1-N2_GFP
FEATURES             Location/Qualifiers
     source          1..5781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     837..856
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     940..959
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        940..958
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             990..1012
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1022..1039
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1040..1060
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2165..2188
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2250..2268)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2264..2311
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2677..2682,2683..2699,2700..2705))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2732..2750)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2818..2922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2923..2991,2992..3783)
                     /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(3141..3160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3954..4542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4443..4462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4696..4713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4728..4867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4793..4812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4953..4975
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4969..5160)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
       61 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
      121 gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt taggctggta
      181 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
      241 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
      301 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
      361 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
      421 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
      481 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
      541 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
      601 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc cttatgcgac
      661 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
      721 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
      781 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
      841 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
      901 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actataggga
      961 caccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     1021 gcaccatcac catcaccatg aaaacctgta cttccagtcc ccttcacagt caggaatgaa
     1081 tcctgccgac ctgagcggac tctctggcaa ggaactggca cgcgcacgcc gcgctgcact
     1141 atccaagcaa gggaaagcag ctgtttctaa taaaacggct agcgtaaacc gtagcactaa
     1201 acaggcggca tcttcgatca atacaaatca ggtgcgctct tctgtaaatg aagtgcccac
     1261 tgattaccaa atggcggatc aattgtgctc tacgattgat catgctgact ttggtaccga
     1321 aagcaatcgc gttagagatc tctgccgtca acgcagagag gcactatcaa ctatcggtaa
     1381 aaaagcggtt aaaaccaacg gcaagccgtc gggtcgcgtt cgaccacagc aatcaggtag
     1441 cggaagcggc ggttctaaag gtgaagaact gttcaccggt gttgttccga tcctggttga
     1501 actggatggt gatgttaacg gccacaaatt ctctgttcgt ggtgaaggtg aaggtgatgc
     1561 aaccaacggt aaactgaccc tgaaattcat ctgcactacc ggtaaactgc cggttccatg
     1621 gccgactctg gtgactaccc tgacctatgg tgttcagtgt ttttctcgtt acccggatca
     1681 catgaagcag catgatttct tcaaatctgc aatgccggaa ggttatgtac aggagcgcac
     1741 catttctttc aaagacgatg gcacctacaa aacccgtgca gaggttaaat ttgaaggtga
     1801 tactctggtg aaccgtattg aactgaaagg cattgatttc aaagaggacg gcaacatcct
     1861 gggccacaaa ctggaatata acttcaactc ccataacgtt tacatcaccg cagacaaaca
     1921 gaagaacggt atcaaagcta acttcaaaat tcgccataac gttgaagacg gtagcgtaca
     1981 gctggcggac cactaccagc agaacactcc gatcggtgat ggtccggttc tgctgccgga
     2041 taaccactac ctgtccaccc agtctaaact gtccaaagac ccgaacgaaa agcgcgacca
     2101 catggtgctg ctggagttcg ttactgcagc aggtatcacg cacggcatgg atgagctcta
     2161 caaatggagt catccacagt ttgaaaagta aagcggttaa gatccggctg ctaacaaagc
     2221 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2281 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     2341 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
     2401 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
     2461 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
     2521 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     2581 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     2641 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     2701 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2761 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2941 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3001 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3061 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3121 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3181 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3241 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3301 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3361 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3421 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3481 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3541 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3601 tggtttattg ctgataaatc tggagccggt gagcgtggct ctcgcggtat cattgcagca
     3661 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3721 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3781 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3841 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3901 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3961 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4021 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4081 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4141 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4201 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4261 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4321 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4381 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4441 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4501 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4561 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4621 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4681 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4741 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     4801 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4861 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4921 ggcatccgct tacagacaag ctgtgaccgc ctccgggagc tgcatgtgtc agaggttttc
     4981 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5041 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5101 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5161 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     5221 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     5281 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     5341 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     5401 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     5461 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     5521 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5581 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     5641 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     5701 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     5761 agtggtgaat ccgttagcga g
//
