LOCUS       sequence-210539-        5932 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 5932)
  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 5932)
  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: NK642
FEATURES             Location/Qualifiers
     source          1..5932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(191..259,260..1051)
                     /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(409..428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1222..1810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1711..1730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1964..1981
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2098..2119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2134..2139,2140..2157,2158..2164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2172..2188
                     /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     2177..2199
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2196..2212
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2196..2212
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5528..5545)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5528..5544)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5537..5559)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5753..5772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5872..5894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(join(5932,1..18))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
       61 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      181 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      241 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      301 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      361 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      421 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      481 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      541 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      601 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      661 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      721 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      781 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      841 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      901 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      961 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1021 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1081 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1141 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1201 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1261 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1321 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1381 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1441 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1501 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1561 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1621 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1681 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1741 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1801 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1861 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1921 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1981 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2041 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2101 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2161 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2221 cgccaagctt gtaacgactg cagtcttgtt gtcgtttgtc gtcgtcgttc tctgtctccc
     2281 acccaacttt tgccaattgc aacagtgact tctgtacaat ttgcaacaga taccaacgtg
     2341 tcgagtcagc acaaatggcg aggaaggttc gcgtgcattg cttgcgcgcc tctttgtcat
     2401 gggcgcacgc tgatttgtgt gtgatttgca ccaaatttcc tgccgacaca acttgggccc
     2461 gtaccacacg acccgtttct ggacccgttt ctggcccaga aacgcgaatg ttgcgaaaga
     2521 tgcgaccacg tgctcgctgt cgacaacaca aactcccgca caatccccat gaaaaaaaaa
     2581 gtgcttgctg ggcttgtgtc tggcctgtgt gcgtggcgtg gccgcgtgtg gcttcgttct
     2641 tgcttctgct cactcagcct gttacacgca gagtggctgc gagaaacaag gagaggggtt
     2701 tgcggcattg gcgcatgtgc atctgccttt ttcattgagt cagtctgcag ctcaagaggt
     2761 tggtttcgtc gacggatatg agtgagcttc tatggatgat gagaggtggt ggtggttgta
     2821 tggttggtgg ttggtggttg gtgcaaggcg agccacgcga agagcgacaa ggatgaggga
     2881 gaggagagga gactaggaaa gaggtgaagg gaagggaagg gcgagcaatg acgacgagat
     2941 aaaagacagc caccaccgca cccgcacaaa agcattgctc gcacgcatgg tcgtgtgtta
     3001 tgtttcacac caccaacata accaatacaa gccatcacca atacaagcca gcgctcaagc
     3061 acacacgcac gcacgcacac acaatacaca cgcacgtgaa acgttcacat tgtccgctca
     3121 caaatgcctt tcccttctcg tcgctttcct tgcctttttt tcttcccctc ctgacttcct
     3181 tcctgccttc cttccttcct ccccttctcc gtccaccaac accatttatt tctctgctct
     3241 ttttcctcgt gccacgtgag cagtcttttc gcgtcgtctt cttctttgac cccgtttacc
     3301 gtcaacacca tcaacaacca gcaacatggt ctccaagggc gaggagacca cgatgggcgt
     3361 catcaagccc gacatgaaga tcaagctcaa gatggagggc aacgtcaacg gccacgcctt
     3421 cgtcatcgag ggcgagggcg agggcaagcc ctacgacggc accaacacca tcaacctcga
     3481 ggtcaaggag ggcgcccccc tccccttctc ctacgacatc ctcaccaccg ccttcgccta
     3541 cggcaaccgc gccttcacca agtaccccga cgacatcccc aactacttca agcagtcctt
     3601 ccccgagggc tactcctggg agcgcaccat gaccttcgag gacaagggca tcgtcaaggt
     3661 caagtccgac atctctatgg aggaggactc cttcatctac gagatccacc tcaagggcga
     3721 gaacttcccc cccaacggcc ccgtcatgca gaagaagacc accggctggg acgcctccac
     3781 cgagcgcatg tacgtccgcg acggcgtcct caagggcgac gtgaagcaca agctcctcct
     3841 cgagggcggc ggccaccacc gcgtcgactt caagaccatc taccgcgcca agaaggccgt
     3901 caagctcccc gactaccact tcgtcgacca ccgcatcgag atcctcaacc acaacaagga
     3961 ctacaacaag gtcaccgtct acgagtccgc cgtcgcccgc aactccaccg acggcatgga
     4021 cgagctgtac aaggcttctg gcggtagcgg tggctccagt ggcgacatag gctttgccaa
     4081 cctccccaat caaatccatc gtgaggtcac gctcaagggg tttgacttca ctttgatggt
     4141 cgttgggcgg tctggattgg gaaaatcgac gtttgtcaac accctgttct ccaccgacat
     4201 ctatgaagag gacaagccct atcccccacc ttgtgctcgc attcccagca caacctctgt
     4261 gcaggagagc gtgacaacgc tcattgagaa cgacgttcag ctcacgctca cgctcgtgga
     4321 cacgccgggc tttggaaacg gcatcaacaa cacagactgc tgggtcccca ttgaggagta
     4381 tgttgacaat gcgttctcca agtacgaggg cgaggagtcg aagatcagcc gcacggatat
     4441 ccgtgattgc cgcgtccact gctgcctcta cttcctcact ccttgcatcc acggcgtccg
     4501 tcccctcgac attgaagtca tgaagcgcct gcacacgaag gtgaacatca tccccgtcat
     4561 tgcaaaggcg gattcgctca cagtggacga gtgccagctc ctcaaggaac gcgtgcgcgc
     4621 cacgctggcg gagcacaaca tcaacgtgtt tgagtttccc gaagatgacg atgatgatga
     4681 agaggtgcaa gacgacgttg caaacgccag gtctgcgtgg ccgtttgctg tatcggcctc
     4741 aacacaggta ctcttcgcag atgataagga aatgcgtggc cgcgcctacc cctggggtta
     4801 cgtcgaagtc gacaatccgc agcactcgga cttcgttggc ctgcgcaacc taatcatcag
     4861 aacgtacatg cagcagctga ttgacacgac gaacaacgtc cactacgagc gtttccgtca
     4921 gcagcgcctc tccagcctct ctcttgacac atccatcagt ggaagcccca aatcggatcc
     4981 caacagccaa aacttcatcg aacagctcga aggacagctc aaacaacacg agaagaagct
     5041 tgtgaagatg gagcgagaga tgacagaagt ctttgagaag cgcgtgcgcg aaaaacagca
     5101 gaaaatcaag acgcaagagg cgcagctgct gaaggagcac gaggagtttg aagaaaagtt
     5161 gaagaagcag caaagggagc ttgagatgca gcgcgagcag tttgagcggg agaagaagga
     5221 gtacgaggcc gaggaggaga agcgaaaggc cgcacaggcg cagtccactg gccggcgcaa
     5281 gaagttcttc taagcgcctc gttggtcatt gccaccttgt tgcctctttt gctgttgtcc
     5341 tcgtgtcgac gcgtgccacg tgcgcttgtt gtgagttgat tctgcttctt gtgcacgtca
     5401 tgccgagccc ttgggccccg caataaaccc actatgccag tgtaactatc cttcccccca
     5461 atgtataact gtgacgatca atgagcggtg cccgtctaga ggatccccgg gtaccgagct
     5521 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5581 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5641 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     5701 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     5761 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5821 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5881 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag gg
//
