LOCUS       sequence-322497-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin resistance and mCherry expression for Salpingoeca rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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: NK802
FEATURES             Location/Qualifiers
     source          1..6293
                     /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"
     primer_bind     complement(4253..4273)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5899..5916)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5899..5915)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5908..5930)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6124..6143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6243..6265
                     /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 tctccaaggg cgaggaggac aacatggcta tcatcaagga
     2941 gttcatgcgc ttcaaggtcc acatggaggg ctccgtcaac ggccacgagt tcgagatcga
     3001 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctca aggtgacgaa
     3061 gggcggcccc ctccccttcg cctgggacat cctctccccc cagttcatgt acggctccaa
     3121 ggcttacgtc aagcaccccg ccgacatccc cgactacctc aagctctcct tccccgaggg
     3181 cttcaagtgg gagcgcgtca tgaacttcga ggacggcggc gtcgtcaccg tcacccagga
     3241 ctcctctctt caggacggcg agttcatcta caaggtcaag ctgcgcggca ccaacttccc
     3301 ctccgacggc cccgtcatgc agaagaagac gatgggttgg gaggcttcct ccgagcgcat
     3361 gtaccccgag gacggcgccc tcaagggcga gatcaagcag cgcctcaagc tcaaggacgg
     3421 cggccactac gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tccagctccc
     3481 cggcgcctac aacgtcaaca tcaagctcga catcacctcc cacaacgagg actacaccat
     3541 cgtcgagcag tacgagcgtg ccgagggccg ccacagcacg ggcggtatgg acgagcttta
     3601 caagtcttaa gagctccccc cagcattatc acgtctactg tgtgtatgtt gtcgctctct
     3661 gtttgacctc tctcttgtct actatgttgc ctctctctct ttcttgattg cacccctttc
     3721 ctcgctacga ctactacgac ttccgctacc ttgactttgt tggcttgttg ccaccttttg
     3781 gtcttgcgtt tcgtctgctc gtcactcttg tctctttttg acgacttgtg tgtgatgacg
     3841 gcgtgaggct gtgactgact gactctcaac aacttccaat aaaaacccct acatcgcacg
     3901 tgactgactg cacatttagt gaagggaagg aaaagaagtg aggtgttttc atgtacatga
     3961 ggcttgtcgt tgctcgggca ccgctcattg atcgtcacag ttatacattg gggggaagga
     4021 tagttacact ggcatagtgg gtttattgcg gggcccaagg gctcggcatg acgtgcacaa
     4081 gaagcagaat caactcacaa caagcgcacg tggcacgcgt cgacacgagg acaacagcaa
     4141 aagaggcaac aaggtggcaa tgaccaacga ggcgcaccgg gcttgcgggt catgcaccag
     4201 gtacgaggac cctcggggac ctccacgtcg gcggtgacgg tgaagccgag acgctcgtag
     4261 aaggggaggt tgcgaggagc ggaggtctcg aggaaagcag ggacaccagc acgctcagcg
     4321 gcctcgacgc cagggaggac gacggcagag ccgagaccct tgccctggtg gtcaggagag
     4381 acgccgacgg tggcgaggaa ccaggcgggc tccttggggc ggtggggggc gaggaggccc
     4441 tccatctgct gctgagcggc gagacgggag ccagacagct cagccatgcg ggggccgatc
     4501 tcagcgaaga cagcaccggc ctcgacagac tcaggggtgg tccagacagc gacagcagca
     4561 ccgtcgtcgg cgacccagac cttcccgatg tcgaggccga cgcgggtgag gaacagctcc
     4621 tgcagctcgg tgacacgctc gatgtggcgg tcagggtcga cggtgtgacg ggtagcaggg
     4681 tagtcagcga aggcagcggc gagggtacgg acagcacggg gcacgtcgtc acgggtagcg
     4741 aggcggacgg taggcttgta ctccgtacca gacatgttgc tggttgttga tggtgttgac
     4801 ggtaaacggg gtcaaagaag aagacgacgc gaaaagactg ctcacgtggc acgaggaaaa
     4861 agagcagaga aataaatggt gttggtggac ggagaagggg aggaaggaag gaaggcagga
     4921 aggaagtcag gaggggaaga aaaaaaggca aggaaagcga cgagaaggga aaggcatttg
     4981 tgagcggaca atgtgaacgt ttcacgtgcg tgtgtattgt gtgtgcgtgc gtgcgtgtgt
     5041 gcttgagcgc tggcttgtat tggtgatggc ttgtattggt tatgttggtg gtgtgaaaca
     5101 taacacacga ccatgcgtgc gagcaatgct tttgtgcggg tgcggtggtg gctgtctttt
     5161 atctcgtcgt cattgctcgc ccttcccttc ccttcacctc tttcctagtc tcctctcctc
     5221 tccctcatcc ttgtcgctct tcgcgtggct cgccttgcac caaccaccaa ccaccaacca
     5281 tacaaccacc accacctctc atcatccata gaagctcact catatccgtc gacgaaacca
     5341 acctcttgag ctgcagactg actcaatgaa aaaggcagat gcacatgcgc caatgccgca
     5401 aacccctctc cttgtttctc gcagccactc tgcgtgtaac aggctgagtg agcagaagca
     5461 agaacgaagc cacacgcggc cacgccacgc acacaggcca gacacaagcc cagcaagcac
     5521 tttttttttc atggggattg tgcgggagtt tgtgttgtcg acagcgagca cgtggtcgca
     5581 tctttcgcaa cattcgcgtt tctgggccag aaacgggtcc agaaacgggt cgtgtggtac
     5641 gggcccaagt tgtgtcggca ggaaatttgg tgcaaatcac acacaaatca gcgtgcgccc
     5701 atgacaaaga ggcgcgcaag caatgcacgc gaaccttcct cgccatttgt gctgactcga
     5761 cacgttggta tctgttgcaa attgtacaga agtcactgtt gcaattggca aaagttgggt
     5821 gggagacaga gaacgacgac gacaaacgac aacaagactg cagtcgttac aggatccccg
     5881 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5941 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     6061 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     6121 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6181 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6241 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
