LOCUS       sequence-322498-        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin resistance and LifeAct-mCherry expression for Salpingoeca 
            rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  AUTHORS   Brunet T, Albert M, Roman W, Coyle MC, Spitzer DC, King N
  TITLE     A flagellate-to-amoeboid switch in the closest living relatives of 
            animals.
  JOURNAL   Elife. 2021 Jan 15;10. pii: 61037. doi: 10.7554/eLife.61037.
  PUBMED    33448265
REFERENCE   2  (bases 1 to 6362)
  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"; journalName: "Elife."; 
            date: "2021-01-15"; volume: "10. pii: 61037. doi"; pages: " 
            10.7554/eLife.61037"
COMMENT     Sequence Label: NK803
FEATURES             Location/Qualifiers
     source          1..6362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2897..2947
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     complement(4322..4342)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5968..5985)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5968..5984)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5977..5999)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6193..6212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6312..6334
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gtaacaaact gccctgcacc tcgtctgcca cctcggttgc
     2281 caccttgtcc tcgttcccca ccaccttgtc ctcgtccgct gccagcatcg cgatcagcgt
     2341 gcttgcgctt ggcgctgctg ctgccggact tcccgttatc ggccatactt tttgtggtgt
     2401 ttccaggcga gatgcgctgt tggcaaagca aaatggcaaa acggcctttg ccacacgcca
     2461 tcctcgcctc acctcgacgt gcggctttcc taaacaggaa acctgacgaa acgaattgag
     2521 cgcattggca cacgtgtgct gtgcgctttg tgtcgctgca tgctatccac cacgcaccac
     2581 ccaatcagtc caactgtgac gtgtcacgcc taggcgaaaa agcggcgaga gcgccaaagc
     2641 aaacgccaaa caaaaaccaa agaagagcaa acgcagcgaa ctgatgtgag acaagggggc
     2701 ggtgcaatgc gtcactgaac gtgcaaaagc caaaaacaac caagaagcag ctgcacaaca
     2761 caacaacaac aacaacaaca acaacacctt gttgtcgcag ctctacgttt ggcattttcc
     2821 tgttgttctc tcttgcgatc agtccgtgtc tcttttctcc cccttttccc agaccacaac
     2881 cacaaaacaa ccagccatgg gtgttgctga ccttattaag aaattcgagt ctatctccaa
     2941 ggaagagggc ggtagcggtg gatctatggt ctccaagggc gaggaggaca acatggctat
     3001 catcaaggag ttcatgcgct tcaaggtcca catggagggc tccgtcaacg gccacgagtt
     3061 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctcaa
     3121 ggtgacgaag ggcggccccc tccccttcgc ctgggacatc ctctcccccc agttcatgta
     3181 cggctccaag gcttacgtca agcaccccgc cgacatcccc gactacctca agctctcctt
     3241 ccccgagggc ttcaagtggg agcgcgtcat gaacttcgag gacggcggcg tcgtcaccgt
     3301 cacccaggac tcctctcttc aggacggcga gttcatctac aaggtcaagc tgcgcggcac
     3361 caacttcccc tccgacggcc ccgtcatgca gaagaagacg atgggttggg aggcttcctc
     3421 cgagcgcatg taccccgagg acggcgccct caagggcgag atcaagcagc gcctcaagct
     3481 caaggacggc ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt
     3541 ccagctcccc ggcgcctaca acgtcaacat caagctcgac atcacctccc acaacgagga
     3601 ctacactatc gttgagcagt atgagcgtgc cgagggtcgt cacagcacgg gcggtatgga
     3661 cgagctttac aagtcttaag agctcccccc agcattatca cgtctactgt gtgtatgttg
     3721 tcgctctctg tttgacctct ctcttgtcta ctatgttgcc tctctctctt tcttgattgc
     3781 acccctttcc tcgctacgac tactacgact tccgctacct tgactttgtt ggcttgttgc
     3841 caccttttgg tcttgcgttt cgtctgctcg tcactcttgt ctctttttga cgacttgtgt
     3901 gtgatgacgg cgtgaggctg tgactgactg actctcaaca acttccaata aaaaccccta
     3961 catcgcacgt gactgactgc acatttagtg aagggaagga aaagaagtga ggtgttttca
     4021 tgtacatgag gcttgtcgtt gctcgggcac cgctcattga tcgtcacagt tatacattgg
     4081 ggggaaggat agttacactg gcatagtggg tttattgcgg ggcccaaggg ctcggcatga
     4141 cgtgcacaag aagcagaatc aactcacaac aagcgcacgt ggcacgcgtc gacacgagga
     4201 caacagcaaa agaggcaaca aggtggcaat gaccaacgag gcgcaccggg cttgcgggtc
     4261 atgcaccagg tacgaggacc ctcggggacc tccacgtcgg cggtgacggt gaagccgaga
     4321 cgctcgtaga aggggaggtt gcgaggagcg gaggtctcga ggaaagcagg gacaccagca
     4381 cgctcagcgg cctcgacgcc agggaggacg acggcagagc cgagaccctt gccctggtgg
     4441 tcaggagaga cgccgacggt ggcgaggaac caggcgggct ccttggggcg gtggggggcg
     4501 aggaggccct ccatctgctg ctgagcggcg agacgggagc cagacagctc agccatgcgg
     4561 gggccgatct cagcgaagac agcaccggcc tcgacagact caggggtggt ccagacagcg
     4621 acagcagcac cgtcgtcggc gacccagacc ttcccgatgt cgaggccgac gcgggtgagg
     4681 aacagctcct gcagctcggt gacacgctcg atgtggcggt cagggtcgac ggtgtgacgg
     4741 gtagcagggt agtcagcgaa ggcagcggcg agggtacgga cagcacgggg cacgtcgtca
     4801 cgggtagcga ggcggacggt aggcttgtac tccgtaccag acatgttgct ggttgttgat
     4861 ggtgttgacg gtaaacgggg tcaaagaaga agacgacgcg aaaagactgc tcacgtggca
     4921 cgaggaaaaa gagcagagaa ataaatggtg ttggtggacg gagaagggga ggaaggaagg
     4981 aaggcaggaa ggaagtcagg aggggaagaa aaaaaggcaa ggaaagcgac gagaagggaa
     5041 aggcatttgt gagcggacaa tgtgaacgtt tcacgtgcgt gtgtattgtg tgtgcgtgcg
     5101 tgcgtgtgtg cttgagcgct ggcttgtatt ggtgatggct tgtattggtt atgttggtgg
     5161 tgtgaaacat aacacacgac catgcgtgcg agcaatgctt ttgtgcgggt gcggtggtgg
     5221 ctgtctttta tctcgtcgtc attgctcgcc cttcccttcc cttcacctct ttcctagtct
     5281 cctctcctct ccctcatcct tgtcgctctt cgcgtggctc gccttgcacc aaccaccaac
     5341 caccaaccat acaaccacca ccacctctca tcatccatag aagctcactc atatccgtcg
     5401 acgaaaccaa cctcttgagc tgcagactga ctcaatgaaa aaggcagatg cacatgcgcc
     5461 aatgccgcaa acccctctcc ttgtttctcg cagccactct gcgtgtaaca ggctgagtga
     5521 gcagaagcaa gaacgaagcc acacgcggcc acgccacgca cacaggccag acacaagccc
     5581 agcaagcact ttttttttca tggggattgt gcgggagttt gtgttgtcga cagcgagcac
     5641 gtggtcgcat ctttcgcaac attcgcgttt ctgggccaga aacgggtcca gaaacgggtc
     5701 gtgtggtacg ggcccaagtt gtgtcggcag gaaatttggt gcaaatcaca cacaaatcag
     5761 cgtgcgccca tgacaaagag gcgcgcaagc aatgcacgcg aaccttcctc gccatttgtg
     5821 ctgactcgac acgttggtat ctgttgcaaa ttgtacagaa gtcactgttg caattggcaa
     5881 aagttgggtg ggagacagag aacgacgacg acaaacgaca acaagactgc agtcgttaca
     5941 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
     6001 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6061 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6121 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     6181 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6241 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6301 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6361 ga
//
