LOCUS       sequence-205750-        6247 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 6247)
  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 6247)
  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: NK619
FEATURES             Location/Qualifiers
     source          1..6247
                     /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"
     primer_bind     complement(5853..5870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5853..5869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5862..5884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6078..6097)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6197..6219
                     /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 tgctcacatc 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 gtaacgactg cagtcttgtt gtcgtttgtc gtcgtcgttc
     2281 tctgtctccc acccaacttt tgccaattgc aacagtgact tctgtacaat ttgcaacaga
     2341 taccaacgtg tcgagtcagc acaaatggcg aggaaggttc gcgtgcattg cttgcgcgcc
     2401 tctttgtcat gggcgcacgc tgatttgtgt gtgatttgca ccaaatttcc tgccgacaca
     2461 acttgggccc gtaccacacg acccgtttct ggacccgttt ctggcccaga aacgcgaatg
     2521 ttgcgaaaga tgcgaccacg tgctcgctgt cgacaacaca aactcccgca caatccccat
     2581 gaaaaaaaaa gtgcttgctg ggcttgtgtc tggcctgtgt gcgtggcgtg gccgcgtgtg
     2641 gcttcgttct tgcttctgct cactcagcct gttacacgca gagtggctgc gagaaacaag
     2701 gagaggggtt tgcggcattg gcgcatgtgc atctgccttt ttcattgagt cagtctgcag
     2761 ctcaagaggt tggtttcgtc gacggatatg agtgagcttc tatggatgat gagaggtggt
     2821 ggtggttgta tggttggtgg ttggtggttg gtgcaaggcg agccacgcga agagcgacaa
     2881 ggatgaggga gaggagagga gactaggaaa gaggtgaagg gaagggaagg gcgagcaatg
     2941 acgacgagat aaaagacagc caccaccgca cccgcacaaa agcattgctc gcacgcatgg
     3001 tcgtgtgtta tgtttcacac caccaacata accaatacaa gccatcacca atacaagcca
     3061 gcgctcaagc acacacgcac gcacgcacac acaatacaca cgcacgtgaa acgttcacat
     3121 tgtccgctca caaatgcctt tcccttctcg tcgctttcct tgcctttttt tcttcccctc
     3181 ctgacttcct tcctgccttc cttccttcct ccccttctcc gtccaccaac accatttatt
     3241 tctctgctct ttttcctcgt gccacgtgag cagtcttttc gcgtcgtctt cttctttgac
     3301 cccgtttacc gtcaacacca tcaacaacca gcaacatgtc tgagggcaag cagcacgttt
     3361 ccattgtcat ctgcggccac gtcgatgccg gcaagtcgac cacgaccggc cgtctcatct
     3421 tcgagctcgg cggcatcccc gagcgtgaga tggagaagct caaggccgag gccgacgctc
     3481 tcggcaaggg ctccttcgcc tttgccttct acatggaccg ccagaaggag gagcgcgagc
     3541 gtggtgtgac cattgcctgc accaccaagg agttcttcac ggagtccaag cactacaccg
     3601 tcattgacgc ccctggccat cgtgacttca tcaagaacat gatcaccggt gcctcccagg
     3661 ccgatgtcgc cctcctcatg gttccctccg acggtaactt caccgccgcc attgccaagg
     3721 gcaaccacaa ggccggtgtc gtccagggcc agacccgcca gcacgccgtc ctcatcaacc
     3781 ttctcggcgt caagcagctc attgttggcg tcaacaagat ggactgcgac gttgccaagt
     3841 actccaagga gcgctacgag gagatccgcg acgagatgcg ccacatgctc atcaagtctg
     3901 gctggaagaa ggcctttgtc atggagtctg tccccgtgct ccccatctcc ggctggatgg
     3961 gcgacaacct catcaccaag tccaagaaca tggactggtg gaagggcgtc gacgtcaagg
     4021 ttgagggcga gaccctccac ctcgagacgc tcaaggaggc tcttgagaag atggtccgcg
     4081 tcccccagcg cgccacggac aagcccatgc gcacccccat ctccggtgtc ttcaagatca
     4141 agggtgtcgg tgatgtcctc accggccgcg tcgagcaggg taccgtcaag cccggcgacg
     4201 aggttgtctt cctccccacc cacactgagg ccaacccctg cactggcaag atcttcaccg
     4261 tcgagatgca ccacaagtct gtcgaggctg ccgccactgg cgacaacgtt ggcctcaacg
     4321 tcaagggtct ggacaagaag aacatgcccc gcgtcggtga tgtcatgatc ctcaagtctg
     4381 acaactccct cggcaagtgc aagagcttcc acgcccaggt ccaggtcatg aaccaccctg
     4441 gtgagctcaa ggccggctac tgccccatcg cctttgtccg cacatcccgc tctgccgtcc
     4501 gcatggccga gatttgctgg aagattggca aggagaccgg tggctccaag gccgagaacc
     4561 ccgccaacct caaggccaac gaggtcgccg aggtcaagtt cgagccccag cagcctttcg
     4621 tcgtcgacac cttcaagtcc tgcgagggtc ttggccgcat tgccatcatg gagggtaaca
     4681 ctgtcgtcat gctcggcaag gtcaccaaga tggagcgcgc ttctggcggt agcggtggct
     4741 ccatggtctc caagggcgag gaggacaaca tggctatcat caaggagttc atgcgcttca
     4801 aggtccacat ggagggctcc gtcaacggcc acgagttcga gatcgagggc gagggcgagg
     4861 gccgccccta cgagggcacc cagaccgcca agctcaaggt gacgaagggc ggccccctcc
     4921 ccttcgcctg ggacatcctc tccccccagt tcatgtacgg ctccaaggct tacgtcaagc
     4981 accccgccga catccccgac tacctcaagc tctccttccc cgagggcttc aagtgggagc
     5041 gcgtcatgaa cttcgaggac ggcggcgtcg tcaccgtcac ccaggactcc tctcttcagg
     5101 acggcgagtt catctacaag gtcaagctgc gcggcaccaa cttcccctcc gacggccccg
     5161 tcatgcagaa gaagacgatg ggttgggagg cttcctccga gcgcatgtac cccgaggacg
     5221 gcgccctcaa gggcgagatc aagcagcgcc tcaagctcaa ggacggcggc cactacgacg
     5281 ccgaggtcaa gaccacctac aaggccaaga agcccgtcca gctccccggc gcctacaacg
     5341 tcaacatcaa gctcgacatc acctcccaca acgaggacta caccatcgtc gagcagtacg
     5401 agcgtgccga gggccgccac agcacgggcg gtatggacga gctttacaag tcttaagagc
     5461 tccccccagc attatcacgt ctactgtgtg tatgttgtcg ctctctgttt gacctctctc
     5521 ttgtctacta tgttgcctct ctctctttct tgattgcacc cctttcctcg ctacgactac
     5581 tacgacttcc gctaccttga ctttgttggc ttgttgccac cttttggtct tgcgtttcgt
     5641 ctgctcgtca ctcttgtctc tttttgacga cttgtgtgtg atgacggcgt gaggctgtga
     5701 ctgactgact ctcaacaact tccaataaaa acccctacat cgcacgtgac tgactgcaca
     5761 tttagtgaag ggaaggaaaa gaagtgaggt gttttcatgt acatgaggct tgtcgttgct
     5821 ctagaggatc cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac
     5881 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5941 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6001 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6061 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     6121 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6181 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6241 cgcgcga
//
