LOCUS       Exported                3481 bp ds-DNA     circular SYN 10-MAR-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3481)
  AUTHORS   Junsu Kang Research Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 4, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3481
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     32..66
                     /label=P2A-BsaI-AGTC-ORF-3-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCccgctacaaacttctctctgctg; added: 2022-08-02"
     primer_bind     32..66
                     /label=P2A-BsaI-AGTC-ORF-2-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCcgctacaaacttctctctgctg; added: 2022-08-02"
     primer_bind     32..66
                     /label=P2A-BsaI-AGTC-ORF-1-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCgctacaaacttctctctgctg; added: 2022-08-02"
     CDS             46..102
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 99"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             103..810
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     103..119
                     /label=kozak-EGFP-BsaI-AGTC-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCgccaccatggtgagcaagggcgagga; added: 
                     2022-08-02"
     primer_bind     complement(145..164)
                     /label=C200 mCherry con -r
                     /note="color: black; sequence: tgcaccttgaagcgcatgaa; added:
                     2022-08-10"
     primer_bind     513..532
                     /label=A874 mCherry -F
                     /note="color: black; sequence: ccccgtaatgcagaagaaga; added:
                     2022-08-10"
     misc_feature    689..711
                     /label=C99-mCherry-con-f
                     /note="color: #a6acb3"
     promoter        complement(818..835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    840..974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(1018..1051)
                     /label=SV40-pA-BsaI-CTCA-r
                     /note="color: black; sequence: 
                     gatggtctctTGAGgaattaaaaaacctcccacacct; added: 2022-08-02"
     CDS             1411..2202
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2295..2883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     2784..2803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     3037..3054
                     /label=A757 L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     terminator      3213..3240
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      3332..3418
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     3352..3371
                     /label=A423 rrnb T1 ter -f
                     /note="color: black; sequence: cgttcaccgacaaacaacag; added:
                     2022-07-25"
     primer_bind     3458..3477
                     /label=C133 pME 5' con -f
                     /note="color: black; sequence: ggatgttttcccagtcacga; added:
                     2022-07-25"
     primer_bind     join(3467..3481,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     join(3481,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccggtctcta gtcccgctac aaacttctct
       61 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctgtgagcaa gggcgaggag
      121 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      181 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      241 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
      301 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      361 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      421 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
      481 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
      541 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
      601 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
      661 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
      721 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
      781 accggcggca tggacgagct gtacaagtaa gctagaacta tagtgagtcg tattacgtag
      841 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      901 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      961 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1021 tgtgggaggt tttttaattc ctcaagagac ctctagacca gccaggacag aaatgcctcg
     1081 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga
     1141 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
     1201 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     1261 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     1321 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     1381 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg
     1441 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     1501 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     1561 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     1621 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     1681 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     1741 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     1801 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     1861 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     1921 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     1981 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     2041 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     2101 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     2161 cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga
     2221 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2401 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     2701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2941 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3001 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3061 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3121 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3181 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3241 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3301 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3361 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3421 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3481 t
//
