LOCUS       sequence-204412-        4688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sat2 for Nourseothricin resistance selection of C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4688)
  AUTHORS   Obinata H, Sugimoto A, Niwa S
  TITLE     Streptothricin acetyl transferase 2 (Sat2): A dominant selection 
            marker for Caenorhabditis elegans genome editing.
  JOURNAL   PLoS One. 2018 May 9;13(5):e0197128. doi: 
            10.1371/journal.pone.0197128. eCollection 2018.
  PUBMED    29742140
REFERENCE   2  (bases 1 to 4688)
  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: "PLoS One.";
            date: "2018-05-9"; volume: "13(5):e0197128. doi"; pages: " 
            10.1371/journal.pone.0197128. eCollection 2018"
COMMENT     Sequence Label: pSN444
FEATURES             Location/Qualifiers
     source          1..4688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2207..2229)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2329..2348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2430..2448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2516..2620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2621..2689,2690..3481)
                     /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(2839..2858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3652..4240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4141..4160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4394..4411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4564..4569,4570..4587,4588..4594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4602..4618
                     /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     4607..4629
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgca tttctaaatt ttagtttaat aaaaaaattt
       61 taaaataata cgagttttga gactaacttt caataaaaac caaaaaattg cttcaaaaaa
      121 gagctcgcgc cgatttacga taaacacagc gcgctacgac actccgaaaa ccgcgccgca
      181 ccattacatt ctccgcccca ccgcacggtt tgcggaaggc gatgaaattg ctgggcctaa
      241 aattttaacc ctttttttcc cgtttttcaa ctgttttccc gattttttcg atattttttt
      301 caggttttta gttagaaaag tatataaaaa ttgcattatt tgtagttttc gctcgtaatt
      361 tgtgatagaa aacggccaat ttgcctagat ttgtgattct accaagtgga atttttggga
      421 aaatatcaat attccacgat ttttattcaa atcataacgt ttcgaaatta tcagcttaaa
      481 taaaactaaa ataaataata aaatactaaa aaccctgaat ttctcggttc tcggttcagc
      541 ttgagttcgt tctatacacg ttatcgattt ttgtcatttc gtttaatcat gggttttgat
      601 tcattttaaa tatgaaattt gatttattcc tcgtttcgaa gttggtttgt cttgtccggc
      661 cggtaaacaa atgttattaa aaaaaataat aaatttcgaa atttttcgta atgttccgca
      721 atcgtgtttt tccgtttatc tcgtcttccc ttccttcaaa tatactattt attttctcaa
      781 ttcacatatt ttcagccagc cctgctcaac aagtggaata aaaatgggcg cgccatgaag
      841 atctccgtca tcccagagca agtcgccgag accctcgacg ccgagaacca cttcatcgtc
      901 cgtgaggtct tcgacgtcca cctctccgac caaggattcg agctctccac ccgttccgtc
      961 tccccatacc gtaaggacta catctccgac gacgactccg acgaggactc cgcctgctac
     1021 ggagccttca tcgaccaaga gctcgtcgga aagatcgagc tcaactccac ctggaacgac
     1081 ctcgcctcca tcgagcacat cgtcgtctcc cacacccacc gtggaaaggg agtcgcccac
     1141 tccctcatcg agttcgccaa gaagtgggcc ctctcccgtc aactcctcgg aatccgtctc
     1201 gagacccaaa ccaacaacgt cccagcctgc aacctctacg ccaagtgcgg attcaccctc
     1261 ggaggaatcg acctcttcac ctacaagacc cgtccacaag tctccaacga gaccgccatg
     1321 tactggtact ggttctccgg agcccaagac gacgcctaag gtaccatggt attgatatct
     1381 gagctccgca tcggccgctg tcatcagatc gccatctcgc gcccgtgcct ctgacttcta
     1441 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     1501 tttgttatta tcaaaaaaac ttcttcttaa tttctttgtt ttttagcttc ttttaagtca
     1561 cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa
     1621 aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc
     1681 tgtgtacact tcttatgttt tttttacttc tgataaattt tttttgaaac atcatagaaa
     1741 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat
     1801 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg
     1861 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc
     1921 tttttggggg tttcccctat tgtttgtcaa gagtttcgag gacggcgttt ttcttgctaa
     1981 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     2041 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     2101 gccttaaatg acagaataca ttcccaatat accaaacata actgtttcct actagtcggc
     2161 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2221 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2281 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2341 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2401 aataccgcat caggcggcct taattaaggg cctcgtgata cgcctatttt tataggttaa
     2461 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2521 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2581 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2641 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2701 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2761 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2821 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2881 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2941 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3001 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3061 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3121 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3181 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3241 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3301 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3361 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3421 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3481 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3781 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4021 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     4081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4261 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4321 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4381 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4441 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4501 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4561 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4621 tcacacagga aacagctggg ggtggggggg gggttggggg tggggtgttg ttggggtgtg
     4681 ttttgtgt
//
