LOCUS       sequence-322499-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin resistance and MRLC-mTFP expression for Salpingoeca 
            rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  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: NK804
FEATURES             Location/Qualifiers
     source          1..6824
                     /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"
     regulatory      3422..3431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4784..4804)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6430..6447)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6430..6446)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6439..6461)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6655..6674)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6774..6796
                     /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 ctgctcgtcg tacaaggaag cgccgcgtgc agcgggccac
     2941 gtccaatgtg tttgccatgt ttgacgaggc acagatccag gagttcaagg aggccttcag
     3001 catgattgac caaaaccaca acggcgtcgt cgacgaggag gatctgcggg agatgtactc
     3061 ctccctcggt caagtgcctg atgacgcaca gatcatgaag atgctgagcg aggcaccaaa
     3121 cccaagcaag ctcaacttca ccatgttcct cacgctcttc ggcgacaaga tgaccgggta
     3181 tgatccagaa gacgtgatca agaacgcatt cgcatgcttc gacccggaag gcacgggcaa
     3241 aatagatgag gacaagctgc gtgagttgct gacgacgatg ggagaccggt tcacagacga
     3301 ggaagtggac gaaatgttca gaggagcgcc cattgacagc gatggaaagt tcgactacag
     3361 agaatacacg cgtgtgctga agcacggcgc aaaggatgag tcggactaca aggaaccggt
     3421 cgccaccatg gtctccaagg gcgaggagac cacgatgggc gtcatcaagc ccgacatgaa
     3481 gatcaagctc aagatggagg gcaacgtcaa cggccacgcc ttcgtcatcg agggcgaggg
     3541 cgagggcaag ccctacgacg gcaccaacac catcaacctc gaggtcaagg agggcgcccc
     3601 cctccccttc tcctacgaca tcctcaccac cgccttcgcc tacggcaacc gcgccttcac
     3661 caagtacccc gacgacatcc ccaactactt caagcagtcc ttccccgagg gctactcctg
     3721 ggagcgcacc atgaccttcg aggacaaggg catcgtcaag gtcaagtccg acatctctat
     3781 ggaggaggac tccttcatct acgagatcca cctcaagggc gagaacttcc cccccaacgg
     3841 ccccgtcatg cagaagaaga ccaccggctg ggacgcctcc accgagcgca tgtacgtccg
     3901 cgacggcgtc ctcaagggcg acgtgaagca caagctcctc ctcgagggcg gcggccacca
     3961 ccgcgtcgac ttcaagacca tctaccgcgc caagaaggcc gtcaagctcc ccgactacca
     4021 cttcgtcgac caccgcatcg agatcctcaa ccacaacaag gactacaaca aggtcaccgt
     4081 ctacgagtcc gccgtcgccc gcaactccac cgacggcatg gacgagctgt acaaggctta
     4141 agagctcccc ccagcattat cacgtctact gtgtgtatgt tgtcgctctc tgtttgacct
     4201 ctctcttgtc tactatgttg cctctctctc tttcttgatt gcaccccttt cctcgctacg
     4261 actactacga cttccgctac cttgactttg ttggcttgtt gccacctttt ggtcttgcgt
     4321 ttcgtctgct cgtcactctt gtctcttttt gacgacttgt gtgtgatgac ggcgtgaggc
     4381 tgtgactgac tgactctcaa caacttccaa taaaaacccc tacatcgcac gtgactgact
     4441 gcacatttag tgaagggaag gaaaagaagt gaggtgtttt catgtacatg aggcttgtcg
     4501 ttgctcgggc accgctcatt gatcgtcaca gttatacatt ggggggaagg atagttacac
     4561 tggcatagtg ggtttattgc ggggcccaag ggctcggcat gacgtgcaca agaagcagaa
     4621 tcaactcaca acaagcgcac gtggcacgcg tcgacacgag gacaacagca aaagaggcaa
     4681 caaggtggca atgaccaacg aggcgcaccg ggcttgcggg tcatgcacca ggtacgagga
     4741 ccctcgggga cctccacgtc ggcggtgacg gtgaagccga gacgctcgta gaaggggagg
     4801 ttgcgaggag cggaggtctc gaggaaagca gggacaccag cacgctcagc ggcctcgacg
     4861 ccagggagga cgacggcaga gccgagaccc ttgccctggt ggtcaggaga gacgccgacg
     4921 gtggcgagga accaggcggg ctccttgggg cggtgggggg cgaggaggcc ctccatctgc
     4981 tgctgagcgg cgagacggga gccagacagc tcagccatgc gggggccgat ctcagcgaag
     5041 acagcaccgg cctcgacaga ctcaggggtg gtccagacag cgacagcagc accgtcgtcg
     5101 gcgacccaga ccttcccgat gtcgaggccg acgcgggtga ggaacagctc ctgcagctcg
     5161 gtgacacgct cgatgtggcg gtcagggtcg acggtgtgac gggtagcagg gtagtcagcg
     5221 aaggcagcgg cgagggtacg gacagcacgg ggcacgtcgt cacgggtagc gaggcggacg
     5281 gtaggcttgt actccgtacc agacatgttg ctggttgttg atggtgttga cggtaaacgg
     5341 ggtcaaagaa gaagacgacg cgaaaagact gctcacgtgg cacgaggaaa aagagcagag
     5401 aaataaatgg tgttggtgga cggagaaggg gaggaaggaa ggaaggcagg aaggaagtca
     5461 ggaggggaag aaaaaaaggc aaggaaagcg acgagaaggg aaaggcattt gtgagcggac
     5521 aatgtgaacg tttcacgtgc gtgtgtattg tgtgtgcgtg cgtgcgtgtg tgcttgagcg
     5581 ctggcttgta ttggtgatgg cttgtattgg ttatgttggt ggtgtgaaac ataacacacg
     5641 accatgcgtg cgagcaatgc ttttgtgcgg gtgcggtggt ggctgtcttt tatctcgtcg
     5701 tcattgctcg cccttccctt cccttcacct ctttcctagt ctcctctcct ctccctcatc
     5761 cttgtcgctc ttcgcgtggc tcgccttgca ccaaccacca accaccaacc atacaaccac
     5821 caccacctct catcatccat agaagctcac tcatatccgt cgacgaaacc aacctcttga
     5881 gctgcagact gactcaatga aaaaggcaga tgcacatgcg ccaatgccgc aaacccctct
     5941 ccttgtttct cgcagccact ctgcgtgtaa caggctgagt gagcagaagc aagaacgaag
     6001 ccacacgcgg ccacgccacg cacacaggcc agacacaagc ccagcaagca cttttttttt
     6061 catggggatt gtgcgggagt ttgtgttgtc gacagcgagc acgtggtcgc atctttcgca
     6121 acattcgcgt ttctgggcca gaaacgggtc cagaaacggg tcgtgtggta cgggcccaag
     6181 ttgtgtcggc aggaaatttg gtgcaaatca cacacaaatc agcgtgcgcc catgacaaag
     6241 aggcgcgcaa gcaatgcacg cgaaccttcc tcgccatttg tgctgactcg acacgttggt
     6301 atctgttgca aattgtacag aagtcactgt tgcaattggc aaaagttggg tgggagacag
     6361 agaacgacga cgacaaacga caacaagact gcagtcgtta caggatcccc gggtaccgag
     6421 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     6481 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     6541 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     6601 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     6661 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6721 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6781 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
