LOCUS       sequence-330646-        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     503..523
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2554..2570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2554..2570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2567..2589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2578..2594
                     /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)."
     promoter        complement(join(2602..2608,2609..2626,2627..2632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2647..2668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2785..2802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2956..3544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3036..3055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3715..4506,4507..4575))
                     /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     4338..4357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4576..4680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4748..4766
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4804..4826)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4926..4945
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5139..5161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5153..5170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5154..5170
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgtctggta cggagtacaa gcctaccgtc cgcctcgcta cccgtgacga cgtgccccgt
       61 gctgtccgta ccctcgccgc tgccttcgct gactaccctg ctacccgtca caccgtcgac
      121 cctgaccgcc acatcgagcg tgtcaccgag ctgcaggagc tgttcctcac ccgcgtcggc
      181 ctcgacatcg ggaaggtctg ggtcgccgac gacggtgctg ctgtcgctgt ctggaccacc
      241 cctgagtctg tcgaggccgg tgctgtcttc gctgagatcg gcccccgcat ggctgagctg
      301 tctggctccc gtctcgccgc tcagcagcag atggagggcc tcctcgcccc ccaccgcccc
      361 aaggagcccg cctggttcct cgccaccgtc ggcgtctctc ctgaccacca gggcaagggt
      421 ctcggctctg ccgtcgtcct ccctggcgtc gaggccgctg agcgtgctgg tgtccctgct
      481 ttcctcgaga cctccgctcc tcgcaacctc cccttctacg agcgtctcgg cttcaccgtc
      541 accgccgacg tggaggtccc cgagggtcct cgtacctggt gcatgacccg caagcccggt
      601 gcgcctcgtt ggtcattgcc accttgttgc ctcttttgct gttgtcctcg tgtcgacgcg
      661 tgccacgtgc gcttgttgtg agttgattct gcttcttgtg cacgtcatgc cgagcccttg
      721 ggccccgcaa taaacccact atgccagtgt aactatcctt ccccccaatg tataactgtg
      781 acgatcaatg agcggtgccc gagcaacgac aagcctcatg tacatgaaaa cacctcactt
      841 cttttccttc ccttcactaa atgtgcagtc agtcacgtgc gatgtagggg tttttattgg
      901 aagttgttga gagtcagtca gtcacagcct cacgccgtca tcacacacaa gtcgtcaaaa
      961 agagacaaga gtgacgagca gacgaaacgc aagaccaaaa ggtggcaaca agccaacaaa
     1021 gtcaaggtag cggaagtcgt agtagtcgta gcgaggaaag gggtgcaatc aagaaagaga
     1081 gagaggcaac atagtagaca agagagaggt caaacagaga gcgacaacat acacacagta
     1141 gacgtgataa tgctgggggg agctcttaag acttgtaaag ctcgtccata ccgcccgtgc
     1201 tgtggcggcc ctcggcacgc tcgtactgct cgacgatggt gtagtcctcg ttgtgggagg
     1261 tgatgtcgag cttgatgttg acgttgtagg cgccggggag ctggacgggc ttcttggcct
     1321 tgtaggtggt cttgacctcg gcgtcgtagt ggccgccgtc cttgagcttg aggcgctgct
     1381 tgatctcgcc cttgagggcg ccgtcctcgg ggtacatgcg ctcggaggaa gcctcccaac
     1441 ccatcgtctt cttctgcatg acggggccgt cggaggggaa gttggtgccg cgcagcttga
     1501 ccttgtagat gaactcgccg tcctgaagag aggagtcctg ggtgacggtg acgacgccgc
     1561 cgtcctcgaa gttcatgacg cgctcccact tgaagccctc ggggaaggag agcttgaggt
     1621 agtcggggat gtcggcgggg tgcttgacgt aagccttgga gccgtacatg aactgggggg
     1681 agaggatgtc ccaggcgaag gggagggggc cgcccttcgt caccttgagc ttggcggtct
     1741 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttga
     1801 cggagccctc catgtggacc ttgaagcgca tgaactcctt gatgatagcc atgttgtcct
     1861 cctcgccctt ggagaccatg gctggttgtt ttgtggttgt ggtctgggaa aagggggaga
     1921 aaagagacac ggactgatcg caagagagaa caacaggaaa atgccaaacg tagagctgcg
     1981 acaacaaggt gttgttgttg ttgttgttgt tgttgtgttg tgcagctgct tcttggttgt
     2041 ttttggcttt tgcacgttca gtgacgcatt gcaccgcccc cttgtctcac atcagttcgc
     2101 tgcgtttgct cttctttggt ttttgtttgg cgtttgcttt ggcgctctcg ccgctttttc
     2161 gcctaggcgt gacacgtcac agttggactg attgggtggt gcgtggtgga tagcatgcag
     2221 cgacacaaag cgcacagcac acgtgtgcca atgcgctcaa ttcgtttcgt caggtttcct
     2281 gtttaggaaa gccgcacgtc gaggtgaggc gaggatggcg tgtggcaaag gccgttttgc
     2341 cattttgctt tgccaacagc gcatctcgcc tggaaacacc acaaaaagta tggccgataa
     2401 cgggaagtcc ggcagcagca gcgccaagcg caagcacgct gatcgcgatg ctggcagcgg
     2461 acgaggacaa ggtggtgggg aacgaggaca aggtggcaac cgaggtggca gacgaggtgc
     2521 agggcagttt gttacaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2581 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2641 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2701 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2761 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2941 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3121 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3361 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3541 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3601 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4261 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4621 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4681 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4741 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4801 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4861 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4921 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4981 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     5041 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5101 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5161 gacggccagt gaattcgagc tcggtacccg gggatcctgt aacgactgca gtcttgttgt
     5221 cgtttgtcgt cgtcgttctc tgtctcccac ccaacttttg ccaattgcaa cagtgacttc
     5281 tgtacaattt gcaacagata ccaacgtgtc gagtcagcac aaatggcgag gaaggttcgc
     5341 gtgcattgct tgcgcgcctc tttgtcatgg gcgcacgctg atttgtgtgt gatttgcacc
     5401 aaatttcctg ccgacacaac ttgggcccgt accacacgac ccgtttctgg acccgtttct
     5461 ggcccagaaa cgcgaatgtt gcgaaagatg cgaccacgtg ctcgctgtcg acaacacaaa
     5521 ctcccgcaca atccccatga aaaaaaaagt gcttgctggg cttgtgtctg gcctgtgtgc
     5581 gtggcgtggc cgcgtgtggc ttcgttcttg cttctgctca ctcagcctgt tacacgcaga
     5641 gtggctgcga gaaacaagga gaggggtttg cggcattggc gcatgtgcat ctgccttttt
     5701 cattgagtca gtctgcagct caagaggttg gtttcgtcga cggatatgag tgagcttcta
     5761 tggatgatga gaggtggtgg tggttgtatg gttggtggtt ggtggttggt gcaaggcgag
     5821 ccacgcgaag agcgacaagg atgagggaga ggagaggaga ctaggaaaga ggtgaaggga
     5881 agggaagggc gagcaatgac gacgagataa aagacagcca ccaccgcacc cgcacaaaag
     5941 cattgctcgc acgcatggtc gtgtgttatg tttcacacca ccaacataac caatacaagc
     6001 catcaccaat acaagccagc gctcaagcac acacgcacgc acgcacacac aatacacacg
     6061 cacgtgaaac gttcacattg tccgctcaca aatgcctttc ccttctcgtc gctttccttg
     6121 cctttttttc ttcccctcct gacttccttc ctgccttcct tccttcctcc ccttctccgt
     6181 ccaccaacac catttatttc tctgctcttt ttcctcgtgc cacgtgagca gtcttttcgc
     6241 gtcgtcttct tctttgaccc cgtttaccgt caacaccatc aacaaccagc aac
//
