LOCUS       sequence-330648-        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"
     regulatory      526..535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1888..1908)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3534..3551)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3534..3550)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3543..3565)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3759..3778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3878..3900
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3938..3956)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4024..4128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4129..4197,4198..4989)
                     /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(4347..4366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5160..5748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5649..5668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5902..5919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6036..6057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6072..6077,6078..6095,6096..6102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6110..6126
                     /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     6115..6137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6134..6150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6134..6150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atggctgctc gtcgtacaag gaagcgccgc gtgcagcggg ccacgtccaa tgtgtttgcc
       61 atgtttgacg aggcacagat ccaggagttc aaggaggcct tcagcatgat tgaccaaaac
      121 cacaacggcg tcgtcgacga ggaggatctg cgggagatgt actcctccct cggtcaagtg
      181 cctgatgacg cacagatcat gaagatgctg agcgaggcac caaacccaag caagctcaac
      241 ttcaccatgt tcctcacgct cttcggcgac aagatgaccg ggtatgatcc agaagacgtg
      301 atcaagaacg cattcgcatg cttcgacccg gaaggcacgg gcaaaataga tgaggacaag
      361 ctgcgtgagt tgctgacgac gatgggagac cggttcacag acgaggaagt ggacgaaatg
      421 ttcagaggag cgcccattga cagcgatgga aagttcgact acagagaata cacgcgtgtg
      481 ctgaagcacg gcgcaaagga tgagtcggac tacaaggaac cggtcgccac catggtctcc
      541 aagggcgagg agaccacgat gggcgtcatc aagcccgaca tgaagatcaa gctcaagatg
      601 gagggcaacg tcaacggcca cgccttcgtc atcgagggcg agggcgaggg caagccctac
      661 gacggcacca acaccatcaa cctcgaggtc aaggagggcg cccccctccc cttctcctac
      721 gacatcctca ccaccgcctt cgcctacggc aaccgcgcct tcaccaagta ccccgacgac
      781 atccccaact acttcaagca gtccttcccc gagggctact cctgggagcg caccatgacc
      841 ttcgaggaca agggcatcgt caaggtcaag tccgacatct ctatggagga ggactccttc
      901 atctacgaga tccacctcaa gggcgagaac ttccccccca acggccccgt catgcagaag
      961 aagaccaccg gctgggacgc ctccaccgag cgcatgtacg tccgcgacgg cgtcctcaag
     1021 ggcgacgtga agcacaagct cctcctcgag ggcggcggcc accaccgcgt cgacttcaag
     1081 accatctacc gcgccaagaa ggccgtcaag ctccccgact accacttcgt cgaccaccgc
     1141 atcgagatcc tcaaccacaa caaggactac aacaaggtca ccgtctacga gtccgccgtc
     1201 gcccgcaact ccaccgacgg catggacgag ctgtacaagg cttaagagct ccccccagca
     1261 ttatcacgtc tactgtgtgt atgttgtcgc tctctgtttg acctctctct tgtctactat
     1321 gttgcctctc tctctttctt gattgcaccc ctttcctcgc tacgactact acgacttccg
     1381 ctaccttgac tttgttggct tgttgccacc ttttggtctt gcgtttcgtc tgctcgtcac
     1441 tcttgtctct ttttgacgac ttgtgtgtga tgacggcgtg aggctgtgac tgactgactc
     1501 tcaacaactt ccaataaaaa cccctacatc gcacgtgact gactgcacat ttagtgaagg
     1561 gaaggaaaag aagtgaggtg ttttcatgta catgaggctt gtcgttgctc gggcaccgct
     1621 cattgatcgt cacagttata cattgggggg aaggatagtt acactggcat agtgggttta
     1681 ttgcggggcc caagggctcg gcatgacgtg cacaagaagc agaatcaact cacaacaagc
     1741 gcacgtggca cgcgtcgaca cgaggacaac agcaaaagag gcaacaaggt ggcaatgacc
     1801 aacgaggcgc accgggcttg cgggtcatgc accaggtacg aggaccctcg gggacctcca
     1861 cgtcggcggt gacggtgaag ccgagacgct cgtagaaggg gaggttgcga ggagcggagg
     1921 tctcgaggaa agcagggaca ccagcacgct cagcggcctc gacgccaggg aggacgacgg
     1981 cagagccgag acccttgccc tggtggtcag gagagacgcc gacggtggcg aggaaccagg
     2041 cgggctcctt ggggcggtgg ggggcgagga ggccctccat ctgctgctga gcggcgagac
     2101 gggagccaga cagctcagcc atgcgggggc cgatctcagc gaagacagca ccggcctcga
     2161 cagactcagg ggtggtccag acagcgacag cagcaccgtc gtcggcgacc cagaccttcc
     2221 cgatgtcgag gccgacgcgg gtgaggaaca gctcctgcag ctcggtgaca cgctcgatgt
     2281 ggcggtcagg gtcgacggtg tgacgggtag cagggtagtc agcgaaggca gcggcgaggg
     2341 tacggacagc acggggcacg tcgtcacggg tagcgaggcg gacggtaggc ttgtactccg
     2401 taccagacat gttgctggtt gttgatggtg ttgacggtaa acggggtcaa agaagaagac
     2461 gacgcgaaaa gactgctcac gtggcacgag gaaaaagagc agagaaataa atggtgttgg
     2521 tggacggaga aggggaggaa ggaaggaagg caggaaggaa gtcaggaggg gaagaaaaaa
     2581 aggcaaggaa agcgacgaga agggaaaggc atttgtgagc ggacaatgtg aacgtttcac
     2641 gtgcgtgtgt attgtgtgtg cgtgcgtgcg tgtgtgcttg agcgctggct tgtattggtg
     2701 atggcttgta ttggttatgt tggtggtgtg aaacataaca cacgaccatg cgtgcgagca
     2761 atgcttttgt gcgggtgcgg tggtggctgt cttttatctc gtcgtcattg ctcgcccttc
     2821 ccttcccttc acctctttcc tagtctcctc tcctctccct catccttgtc gctcttcgcg
     2881 tggctcgcct tgcaccaacc accaaccacc aaccatacaa ccaccaccac ctctcatcat
     2941 ccatagaagc tcactcatat ccgtcgacga aaccaacctc ttgagctgca gactgactca
     3001 atgaaaaagg cagatgcaca tgcgccaatg ccgcaaaccc ctctccttgt ttctcgcagc
     3061 cactctgcgt gtaacaggct gagtgagcag aagcaagaac gaagccacac gcggccacgc
     3121 cacgcacaca ggccagacac aagcccagca agcacttttt ttttcatggg gattgtgcgg
     3181 gagtttgtgt tgtcgacagc gagcacgtgg tcgcatcttt cgcaacattc gcgtttctgg
     3241 gccagaaacg ggtccagaaa cgggtcgtgt ggtacgggcc caagttgtgt cggcaggaaa
     3301 tttggtgcaa atcacacaca aatcagcgtg cgcccatgac aaagaggcgc gcaagcaatg
     3361 cacgcgaacc ttcctcgcca tttgtgctga ctcgacacgt tggtatctgt tgcaaattgt
     3421 acagaagtca ctgttgcaat tggcaaaagt tgggtgggag acagagaacg acgacgacaa
     3481 acgacaacaa gactgcagtc gttacaggat ccccgggtac cgagctcgaa ttcactggcc
     3541 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3601 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3661 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3721 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3781 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3841 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3901 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3961 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4021 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4081 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4141 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4201 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4261 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4321 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4381 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4441 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4501 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4561 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4621 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4681 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4741 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4801 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4861 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4921 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4981 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5041 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5101 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5161 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5221 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5281 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5341 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5401 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5461 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5521 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5581 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5641 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5701 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5761 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5821 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5881 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5941 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6001 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6061 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6121 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgt aacaaactgc
     6181 cctgcacctc gtctgccacc tcggttgcca ccttgtcctc gttccccacc accttgtcct
     6241 cgtccgctgc cagcatcgcg atcagcgtgc ttgcgcttgg cgctgctgct gccggacttc
     6301 ccgttatcgg ccatactttt tgtggtgttt ccaggcgaga tgcgctgttg gcaaagcaaa
     6361 atggcaaaac ggcctttgcc acacgccatc ctcgcctcac ctcgacgtgc ggctttccta
     6421 aacaggaaac ctgacgaaac gaattgagcg cattggcaca cgtgtgctgt gcgctttgtg
     6481 tcgctgcatg ctatccacca cgcaccaccc aatcagtcca actgtgacgt gtcacgccta
     6541 ggcgaaaaag cggcgagagc gccaaagcaa acgccaaaca aaaaccaaag aagagcaaac
     6601 gcagcgaact gatgtgagac aagggggcgg tgcaatgcgt cactgaacgt gcaaaagcca
     6661 aaaacaacca agaagcagct gcacaacaca acaacaacaa caacaacaac aacaccttgt
     6721 tgtcgcagct ctacgtttgg cattttcctg ttgttctctc ttgcgatcag tccgtgtctc
     6781 ttttctcccc cttttcccag accacaacca caaaacaacc agcc
//
