LOCUS       sequence-485235-        5933 bp    DNA     circular SYN 03-SEP-2025
DEFINITION  Bacterial expression of SynPspA with C-terminal YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5933)
  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 5933)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 3, 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: pBAD_SynPspA
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(1993..1995,1996..2709)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2035..2056)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2296..2315)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2643..2664
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2791..2808)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2961..3047
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3139..3166
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3185..3276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3277..3345,3346..4137)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTATNLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIVDKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3495..3514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4179..4634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4266..4285)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4476..4497
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4745..5333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5234..5253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5487..5504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5670..5692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5830..5849
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcaatg
     1321 gaactattta atagggttgg aagagtattg aagtcccaat tgacccattg gcagcagcag
     1381 caagaagctc cggaagatct gttggagcgc ctgctgggcg aaatggaatt ggagctgatc
     1441 gagctgcgtc gcgcactggc gcagaccatt gcgacgttta aaagcactga gcgccaacgt
     1501 gatgcgcaac agctgattgc ccaacgttgg tacgaaaagg cgcaggcggc gttggaccgt
     1561 ggtaatgagc aactagctcg tgaagcgctg ggtcagcgtc agagctatca gtcccacacc
     1621 gaagcgctcg gcaaatctct gggtgaacaa cgtgcattgg tcgagcaggt tagaggccag
     1681 ctgcaaaagc tggagcgtaa atacctggag cttaagagcc agaagaacct ttatctggcc
     1741 cgcctgaagt cagcaattgc tgcgcagaag atcgaggaga tcgcgggtaa cctggacaac
     1801 gcgagcgcaa gctctctgtt cgaacggatt gaaaccaaaa tcctggagtt agaggccgaa
     1861 cgtgaactgt tgaatccgcc tccgtccccg ctggacaaaa aattcgagca gtgggaagag
     1921 caacaagctg tggaagcaac cctggctgcc atgaaagcgc gccgtagctt accgccgcca
     1981 agctcgggat ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2041 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2101 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     2161 tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac
     2221 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2281 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2341 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2401 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     2461 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     2521 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2581 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2641 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2701 tacaagtaac catggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
     2761 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     2821 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     2881 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     2941 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3001 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     3061 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     3121 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     3181 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3301 gcccttattc ccttttttgc ggcattttgt cttcctgttt ttgctcaccc agaaacgctg
     3361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3481 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     3541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3661 gataacactg cgaccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3961 attgttgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4141 tcagaccaag tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt
     4201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4381 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4501 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     4561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4621 aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4861 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5521 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtcatggc
     5581 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5641 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5701 gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc cggccacggg
     5761 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5821 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5881 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atc
//
