LOCUS       sequence-210416-        4601 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 4601)
  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 4601)
  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: NK585
FEATURES             Location/Qualifiers
     source          1..4601
                     /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     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2143..2148,2149..2166,2167..2173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2197
                     /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     2186..2208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2205..2221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2205..2221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4206..4223)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4206..4222)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4215..4237)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4431..4450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4550..4572
                     /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 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2041 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2101 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2161 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2221 ccatgattac gccaagcttg taagtagtgg tagtcgtcga agacgtgggg tgcggggatg
     2281 tgcttgactt ggtgggagag gctggtggaa gtagctgatg aaaatgaact ccaccacacg
     2341 ccaactgcca cttgaaccac ggaaggcagg cagcagtgct gctgtgtgcg cttttcggcg
     2401 cggcgcgcag ccacaaagca acggcaattg tggcgggtga ttcgttcgaa cgctgctttt
     2461 cataattttc gccaatgtct gaatcaatgt ctttttcaag ggtgcagaat aacgaacatt
     2521 attttttatt gtgtggattc agcgccaatt gcgatttgga gatgcgctaa gcgcacaatc
     2581 accaccaaac acgacattcc ccgctttccc gccaaactga ttctcgcgcg aaacctgaga
     2641 tgtgcaccgc gccgttacaa gtgcgaccac gcacacctct tcttgtcatt tatcatccgc
     2701 atctccaagc aagcaagcaa gcaaaccagg accccctccc tcgctcgtca cctcaccacc
     2761 tcccaaactt aaacttcaca accagcacaa tggcccgcac gaagcagacc gcccgcaaga
     2821 ccaccggttc tggtggctct ggtggcatgg tcttcaccct cgaggacttc gtcggcgact
     2881 ggcgccagac cgccggctac aacctcgacc aggtcctcga gcagggcggc gtctcctccc
     2941 tcttccagaa cctcggcgtc tccgtcaccc ccatccagcg catcgtcctc tccggcgaga
     3001 acggcctcaa gatcgacatc cacgtcatca tcccctacga gggcctctcc ggcgaccaga
     3061 tgggccagat cgagaagatc ttcaaggtcg tctaccccgt cgacgaccac cacttcaagg
     3121 tcatcctcca ctacggcacc ctcgtcatcg acggcgtcac ccccaacatg atcgactact
     3181 tcggccgccc ctacgagggc atcgccgtct tcgacggcaa gaagatcacc gtcaccggca
     3241 ccctctggaa cggcaacaag atcatcgacg agcgcctcat caaccccgac ggctccctcc
     3301 tcttccgcgt caccatcaac ggcgtcaccg gctggcgcct ctgcgagcgc atcctcgccg
     3361 cttaagcgtc tcaccttctc aacatggctc gctcgctcga tgtcgttctc tgacaccttg
     3421 cttggcttgt cgtcatgagg ttgctttgtc atcaccaacg ctggttgcat tttgatctgt
     3481 gacaccttga atcatcctcc ctcccctccc ttgttcttaa ttacccactt ggcccttgct
     3541 gtctctcttg gtctgtgcca cgggttgtcg cctccgcggg ctcaaattgt gctcacacgt
     3601 cttttatcca tgctcgtgtt cctgagcact tgacgctggc ggtgtctttg tgcgctcgtg
     3661 tgcatgggaa cctctctccc tccctctccc tgtgtgtaat ggcgttggct taccgtggta
     3721 tcgatttgcc tctgcgtaat gacacgccgc aacggcgttt ggggagtttc cccggcttgt
     3781 cctgcctcgc aacccttctc ctttggttcg ttgttgcctc tcaatacaat gatcaaaccg
     3841 aacattaatg tgaggaccat gatggcatcc tgctttattc aagcaagtgg aatgaatgac
     3901 acagaaacag attgatcgtc attgtgatgc cgtgcgtgta tttgtggcgg agaggaacaa
     3961 aagtcacagg tggctacatt tacacaactt cttgtcaatg caaactcagt gagtcattgg
     4021 gtatcatgcg tggtgaagaa tgagggagtg ctgccagcat ctgaacgagc accatcgtgc
     4081 cagggcgcgg ctcttggcaa actcgcgtgc gtccatctct tcgtgatcat cgtcatcatc
     4141 gcgacgctgt gtcacatgtg ggacaatgaa tgtctagagg atccccgggt accgagctcg
     4201 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4261 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4321 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     4381 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     4441 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4501 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4561 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga g
//
