LOCUS       sequence-452323-        3489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of SynPspA with N-terminal 10xHis-tag and EK 
            cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3489)
  AUTHORS   Junglas B, Huber ST, Heidler T, Schlosser L, Mann D, Hennig R, 
            Clarke M, Hellmann N, Schneider D, Sachse C
  TITLE     PspA adopts an ESCRT-III-like fold and remodels bacterial membranes.
  JOURNAL   Cell. 2021 Jul 8;184(14):3674-3688.e18. doi: 
            10.1016/j.cell.2021.05.042. Epub 2021 Jun 23.
  PUBMED    34166616
REFERENCE   2  (bases 1 to 3489)
  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: 
            "10.1016/j.cell.2021.05.042"; journalName: "Cell"; date: "2021-07-8-
            8"; volume: "184"; issue: "14"; pages: "3674-3688.e18"
COMMENT     Sequence Label: pRSET_SynPspA
FEATURES             Location/Qualifiers
     source          1..3489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(164..183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      977..1565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1466..1485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1719..1736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1821..1840
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1821..1839
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1871..1893
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1906..1935
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1951..1965
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(2691..2709)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2705..2752
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2849..3304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2936..2955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3146..3167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3330..3434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3435..3489,1..14,15..806)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
       61 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      181 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1681 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1741 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1801 ggatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctm
     1861 tagaaataat tttgtttaac tttaagaagg agatatacca tgggccatca tcatcatcat
     1921 catcatcatc atcacagcag cggccatatc gacgacgacg acaagcatat ggaacttttt
     1981 aatcgggttg gacgggtgtt gaaatcccag ttaacccatt ggcaacagca acaggaagcg
     2041 ccggaggatc tcctcgaacg tctattgggc gagatggagt tggagctgat cgaattgcgg
     2101 cgggccttgg cccaaaccat tgctactttt aaaagtacgg aacgacagag agatgctcag
     2161 cagttaattg cccaacgctg gtacgaaaag gctcaggctg ccctggatag gggtaatgag
     2221 caattggcta gggaggccct tgggcaaaga cagtcctatc agagtcatac ggaagcccta
     2281 ggtaaatccc tcggggaaca acgggccctg gtggaacagg tgcggggaca attacaaaag
     2341 ctggagcgca aatatctgga gctgaaaagt cagaaaaatc tttacctcgc ccgcttaaaa
     2401 tcggcgatcg ccgcccaaaa aattgaagag attgcgggca atttggacaa tgccagtgcc
     2461 agtagtttat ttgagagaat tgaaaccaaa atcctggagt tggaagcgga acgggaattg
     2521 ctcaaccctc ccccatcccc gctagataaa aaatttgagc agtgggagga acaacaggcg
     2581 gtggaagcaa cccttgcggc catgaaagct cgacgctctc tccctccccc ttcctcgtaa
     2641 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2701 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2761 aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2821 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2881 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2941 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3001 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3061 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3121 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3181 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3241 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3301 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3481 cattttgcc
//
