LOCUS       sequence-210270-        4684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in S. rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4684)
  AUTHORS   Booth DS, Szmidt-Middleton H, King N
  TITLE     Transfection of choanoflagellates illuminates their cell biology and
            the ancestry of animal septins.
  JOURNAL   Mol Biol Cell. 2018 Dec 1;29(25):3026-3038. doi: 
            10.1091/mbc.E18-08-0514. Epub 2018 Oct 3.
  PUBMED    30281390
REFERENCE   2  (bases 1 to 4684)
  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"; doi: 
            "10.1091/mbc.E18-08-0514"; journalName: "Mol Biol Cell"; date: 
            "2018-12-1- 1"; volume: "29"; issue: "25"; pages: "3026-3038"
COMMENT     Sequence Label: NK606
FEATURES             Location/Qualifiers
     source          1..4684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        95..199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(200..268,269..1060)
                     /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(418..437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1231..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2012..2029
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2146..2167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2182..2187,2188..2205,2206..2212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2220..2236
                     /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     2225..2247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2244..2260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2244..2260
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4289..4306)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4289..4305)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4298..4320)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4514..4533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4633..4655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      121 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      181 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      241 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      301 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      361 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      421 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      481 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      541 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      601 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      661 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      721 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      781 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      841 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      901 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      961 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1021 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1081 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1141 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1201 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1261 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1321 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1381 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatct tctttcctgc gttatcccgc tggccttttg ctcacatctt
     1921 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1981 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2041 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2101 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     2161 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     2221 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgt
     2281 aacgactgca gtcttgttgt cgtttgtcgt cgtcgttctc tgtctcccac ccaacttttg
     2341 ccaattgcaa cagtgacttc tgtacaattt gcaacagata ccaacgtgtc gagtcagcac
     2401 aaatggcgag gaaggttcgc gtgcattgct tgcgcgcctc tttgtcatgg gcgcacgctg
     2461 atttgtgtgt gatttgcacc aaatttcctg ccgacacaac ttgggcccgt accacacgac
     2521 ccgtttctgg acccgtttct ggcccagaaa cgcgaatgtt gcgaaagatg cgaccacgtg
     2581 ctcgctgtcg acaacacaaa ctcccgcaca atccccatga aaaaaaaagt gcttgctggg
     2641 cttgtgtctg gcctgtgtgc gtggcgtggc cgcgtgtggc ttcgttcttg cttctgctca
     2701 ctcagcctgt tacacgcaga gtggctgcga gaaacaagga gaggggtttg cggcattggc
     2761 gcatgtgcat ctgccttttt cattgagtca gtctgcagct caagaggttg gtttcgtcga
     2821 cggatatgag tgagcttcta tggatgatga gaggtggtgg tggttgtatg gttggtggtt
     2881 ggtggttggt gcaaggcgag ccacgcgaag agcgacaagg atgagggaga ggagaggaga
     2941 ctaggaaaga ggtgaaggga agggaagggc gagcaatgac gacgagataa aagacagcca
     3001 ccaccgcacc cgcacaaaag cattgctcgc acgcatggtc gtgtgttatg tttcacacca
     3061 ccaacataac caatacaagc catcaccaat acaagccagc gctcaagcac acacgcacgc
     3121 acgcacacac aatacacacg cacgtgaaac gttcacattg tccgctcaca aatgcctttc
     3181 ccttctcgtc gctttccttg cctttttttc ttcccctcct gacttccttc ctgccttcct
     3241 tccttcctcc ccttctccgt ccaccaacac catttatttc tctgctcttt ttcctcgtgc
     3301 cacgtgagca gtcttttcgc gtcgtcttct tctttgaccc cgtttaccgt caacaccatc
     3361 aacaaccagc aacatgtctg tcttcaccct cgaggacttc gtcggcgact ggcgccagac
     3421 cgctggttac aacctcgacc aggtcctcga gcagggcggc gtctcctccc tcttccagaa
     3481 cctcggcgtc tccgtcaccc ccatccagcg catcgtcctc tccggcgaga acggcctcaa
     3541 gatcgacatc cacgtcatca tcccctacga gggcctctcc ggcgaccaga tgggccagat
     3601 cgagaagatc ttcaaggtcg tctaccccgt cgacgaccac cacttcaagg tcatcctcca
     3661 ctacggcacc ctcgtcatcg acggcgtcac ccccaacatg atcgactact tcggccgccc
     3721 ctacgagggc atcgccgtct tcgacggcaa gaagatcacc gtcaccggca ccctctggaa
     3781 cggcaacaag atcatcgacg agcgcctcat caaccccgac ggctccctcc tcttccgcgt
     3841 caccatcaac ggcgtcaccg gctggcgcct ctgcgagcgc atcctcgctt aagagctccc
     3901 cccagcatta tcacgtctac tgtgtgtatg ttgtcgctct ctgtttgacc tctctcttgt
     3961 ctactatgtt gcctctctct ctttcttgat tgcacccctt tcctcgctac gactactacg
     4021 acttccgcta ccttgacttt gttggcttgt tgccaccttt tggtcttgcg tttcgtctgc
     4081 tcgtcactct tgtctctttt tgacgacttg tgtgtgatga cggcgtgagg ctgtgactga
     4141 ctgactctca acaacttcca ataaaaaccc ctacatcgca cgtgactgac tgcacattta
     4201 gtgaagggaa ggaaaagaag tgaggtgttt tcatgtacat gaggcttgtc gttgctctag
     4261 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
     4321 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4381 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4441 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4501 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4561 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4621 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4681 cgag
//
