LOCUS       Exported                3371 bp ds-DNA     circular SYN 13-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3371)
  AUTHORS   Broad Institute
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3371
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    432..443
                     /label=GG-5F
     CDS             438..443
                     /codon_start=1
                     /label=GS
                     /translation="GS"
     CDS             444..500
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A_v1.1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             501..810
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferasecontains 304C>A 
                     silent mutation"
                     /label=SplitPuroN_v1.1
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA"
     misc_feature    808..827
                     /label=CBE_inducible_splice_donor_target_v2 [Sp]
     intron          828..907
                     /label=modSV40_intron_v2
                     /note="SV40 intron modified to contain a programmed sgRNA 
                     target, PAM sequence and stop codonsequence originally from
                     Xu DH et al, J Cell Mol Med. 2018"
     misc_difference 834..836
                     /label=STOP in frame
     CDS             908..1191
                     /codon_start=3
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=SplitPuro-C_v1.1
                     /note="confers resistance to puromycin"
                     /translation="QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEA
                     AERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             1192..1197
                     /label=GS
     CDS             1198..1200
                     /label=Stop
     misc_feature    1204..1215
                     /label=GG-6R
     rep_origin      complement(1683..2271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2449..3258)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgga agcgcaacaa acttctctct gctgaaacaa gccggagatg
      481 tcgaagagaa tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
      541 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
      601 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
      661 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
      721 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
      781 tggccgagtt gagcggttcc cggctggcag gcgtgtaact tgaacggggg ttttaattca
      841 ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat gttgccttta
      901 cttctagcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
      961 tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     1021 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1081 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1141 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc cggatcctga
     1201 taaattccag tcttcggaac caaatcggat cccgggcccg tcgactgcag aggcctgcat
     1261 gcaagcttgg tgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1321 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1381 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1441 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1501 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1561 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1621 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2221 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2281 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2341 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2401 aaatcaagcc caatctgaat aatgttacaa ccaattaacc aattctgatt agaaaaactc
     2461 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     2521 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     2581 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2641 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2701 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2761 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2821 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2881 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2941 ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     3001 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     3061 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     3121 atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     3181 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc
     3241 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     3301 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg
     3361 ccagagctgc a
//
