LOCUS       Exported                3008 bp ds-DNA     circular SYN 20-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3008)
  AUTHORS   Broad Institute
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 30, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3008
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2909..2983
                     /organism="Spizellomyces punctatus DAOM BR117"
                     /chromosome="Unknown"
                     /strain="DAOM BR117"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:645134"
     misc_feature    1..30
                     /label=Overhung
     primer_bind     1..30
                     /label=Vector.REV
     primer_bind     1..22
                     /label=Vector.FOR
     promoter        14..343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      194..329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(235..256)
                     /label=Primer 738
     primer_bind     complement(329..348)
                     /label=Primer 1
     polyA_signal    349..470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     349..373
                     /label=Primer 2
     primer_bind     complement(519..535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    543..559
                     /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)."
     promoter        complement(567..597)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(921..1506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1677..2537)
                     /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     2427..2447
                     /label=Primer 261
     promoter        complement(2538..2642)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2643..2667
                     /label=Overhung
     primer_bind     complement(2645..2667)
                     /label=Vector.FOR
     promoter        2668..2908
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2668..2688
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2839..2858
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2879..2908
                     /label=Overhung
     primer_bind     complement(2883..2908)
                     /label=Vector.REV
     ncRNA           2909..2983
                     /label=ncRNA scaffold
                     /ncRNA_class="other"
     misc_feature    2909..2983
                     /label=Spu-v3
     misc_feature    complement(2951..2983)
                     /standard_name="TAGTGCGGAGCCGATGGGTATTGGCACTAGTAC"
                     /label=TAGTGCGGAGCCGATGGGTATTGGCACTAGTAC
                     /note="Geneious type: motif3"
     misc_feature    complement(2951..2983)
                     /standard_name="TAGTGCGGAGCCAATGGGTATTGGCACTAGCAC"
                     /label=TAGTGCGGAGCCAATGGGTATTGGCACTAGCAC
                     /note="Geneious type: motif2"
     misc_feature    2953
                     /standard_name="T"
                     /label=T
                     /note="Geneious type: mismatch"
     misc_feature    2970..2971
                     /standard_name="AA"
                     /label=AA
                     /note="Geneious type: mismatch"
     misc_feature    2970
                     /standard_name="A"
                     /label=A
                     /note="Geneious type: mismatch"
     misc_feature    2986..2991
                     /label=BbsI
     misc_feature    2995..3000
                     /label=BbsI
     misc_feature    3003..3008
                     /label=U6 terminator
ORIGIN
        1 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
       61 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
      181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
      241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
      301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
      361 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      421 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
      481 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
      541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1081 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1321 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1441 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2521 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2641 cgcacatttc cccgaaaagt gccacctgag ggcctatttc ccatgattcc ttcatatttg
     2701 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2761 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2821 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2881 ttggctttat atatcttgtg gaaaggacgt tttccgagcc ggttgtcgcg cggttcaatc
     2941 cctggtgcgg gtgctagtgc caatacccac cggctccgca ctacagtctt ctctgaagac
     3001 catttttt
//
