LOCUS       sequence-297998-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged second N-peptide of NTD fused to Superfolder GFP 
            ([N2]-GFP) used for BLI, pET-14b backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  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: pBz173_CsoS2_N2_GFP
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      528..1116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1017..1036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1270..1287
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1302..1441
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1367..1386)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1527..1549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1543..1734)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3192..3211
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3295..3314
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3295..3313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3345..3367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3380..3397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3818..3838
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4571..4594
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4656..4674)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4670..4717
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5083..5088,5089..5105,5106..5111))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5138..5156)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5224..5328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5329..5397,5398..5832,1..357)
                     /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(5547..5566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
       61 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      121 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      181 attgctgata aatctggagc cggtgagcgt ggctctcgcg gtatcattgc agcactgggg
      241 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      301 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
      361 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      421 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      481 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      661 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1141 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1201 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1261 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     1321 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     1381 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     1441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     1501 cgcttacaga caagctgtga ccgcctccgg gagctgcatg tgtcagaggt tttcaccgtc
     1561 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     1621 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     1681 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     1741 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     1801 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     1861 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     1921 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     1981 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     2041 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     2101 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     2161 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     2221 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     2281 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     2341 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     2401 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     2461 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     2521 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     2581 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     2641 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     2701 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     2761 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     2821 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     2881 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     2941 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgacgc
     3001 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     3061 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     3121 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     3181 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     3241 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     3301 gactcactat agggacacca caacggtttc cctctagaaa taattttgtt taactttaag
     3361 aaggagatat accatgtccc accatcatca tcatcatgca gctgtttcta ataaaacggc
     3421 tagcgtaaac cgtagcacta aacaggcggc atcttcgatc aatacaaatc aggtgcgctc
     3481 ttctgtaaat gaagtgccca ctgattacca aatggcggat caattgtgct ctacgattga
     3541 tcatgctgac tttggtaccg aaagcaatcg cgttagagat ctctgccgtc aacgcagaga
     3601 ggcactatca actatcggta aaaaagcggt taaaaccaac ggcaagccgt cgggtcgcgt
     3661 tcgaccacag caatcagtgg ttcacaacga cgcaatgatc gaaaatgccg gtgatactaa
     3721 ccaatcatcg tccacttcat taaataatga actttccgaa atctgctcca tagcagacga
     3781 catgccggag cgttttggtt cacaagccaa aaccgtcgaa aacctgtact tccagtcctc
     3841 aggtagcgga agcggcggtt ctaaaggtga agaactgttc accggtgttg ttccgatcct
     3901 ggttgaactg gatggtgatg ttaacggcca caaattctct gttcgtggtg aaggtgaagg
     3961 tgatgcaacc aacggtaaac tgaccctgaa attcatctgc actaccggta aactgccggt
     4021 tccatggccg actctggtga ctaccctgac ctatggtgtt cagtgttttt ctcgttaccc
     4081 ggatcacatg aagcagcatg atttcttcaa atctgcaatg ccggaaggtt atgtacagga
     4141 gcgcaccatt tctttcaaag acgatggcac ctacaaaacc cgtgcagagg ttaaatttga
     4201 aggtgatact ctggtgaacc gtattgaact gaaaggcatt gatttcaaag aggacggcaa
     4261 catcctgggc cacaaactgg aatataactt caactcccat aacgtttaca tcaccgcaga
     4321 caaacagaag aacggtatca aagctaactt caaaattcgc cataacgttg aagacggtag
     4381 cgtacagctg gcggaccact accagcagaa cactccgatc ggtgatggtc cggttctgct
     4441 gccggataac cactacctgt ccacccagtc taaactgtcc aaagacccga acgaaaagcg
     4501 cgaccacatg gtgctgctgg agttcgttac tgcagcaggt atcacgcacg gcatggatga
     4561 gctctacaaa tggagtcatc cacagtttga aaagtaaagc ggttaagatc cggctgctaa
     4621 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     4681 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     4741 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     4801 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     4861 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     4921 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     4981 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     5041 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     5101 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5161 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5221 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5281 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5341 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5821 ccggagctga at
//
