LOCUS       Exported                3474 bp ds-DNA     circular SYN 16-SEP-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 4, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3474
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..87
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     21..40
                     /label=A423 rrnb T1 ter -f
                     /note="color: black; sequence: cgttcaccgacaaacaacag; added:
                     2022-07-25"
     primer_bind     127..146
                     /label=C133 pME 5' con -f
                     /note="color: black; sequence: ggatgttttcccagtcacga; added:
                     2022-07-25"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     182..215
                     /label=P2A-BsaI-AGTC-ORF-3-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCccgctacaaacttctctctgctg; added: 2022-08-02"
     primer_bind     182..215
                     /label=P2A-BsaI-AGTC-ORF-2-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCcgctacaaacttctctctgctg; added: 2022-08-02"
     primer_bind     182..215
                     /label=P2A-BsaI-AGTC-ORF-1-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCgctacaaacttctctctgctg; added: 2022-08-02"
     CDS             195..251
                     /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 249"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(227..269)
                     /label=2a-BFP-noATG-vec-r
                     /note="color: black; sequence: 
                     CTCCTTAATCAGCTCGCTaggtccagggttctcctccacgtct; added: 
                     2022-08-16"
     primer_bind     230..273
                     /label=2a-BFP-noATG-ins-f
                     /note="color: black; sequence: 
                     cgtggaggagaaccctggacctAGCGAGCTGATTAAGGAGAACA; added: 
                     2022-08-16"
     CDS             252..950
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /note="color: #3366ff"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     252..270
                     /label=BFP-LS-ins-f
                     /note="color: black; sequence: 
                     cccggtctctAGTCgccaccATGAGCGAGCTGATTAAGGAGA; added: 
                     2022-08-16"
     primer_bind     252..269
                     /label=C21 evrogen EcoRI -f
                     /note="color: black; sequence: atgagcgagctgattaaggag"
     primer_bind     complement(358..377)
                     /label=A727 tagBFP-N-r
                     /note="color: black; sequence: gattctcatggtctgggtgc; added:
                     2022-06-21"
     primer_bind     complement(846..864)
                     /label=A898 BFP BsaI -r
                     /note="color: black; sequence: TGGCCTCCTTGATTCTTTC; added: 
                     2022-07-08"
     primer_bind     complement(853..889)
                     /label=BFP BsaI mut -r
                     /note="color: black; sequence: 
                     TGCTGCTCGACGTAcGTCTCGTTGTTGGCCTCCTTGA; added: 2022-07-08"
     primer_bind     854..873
                     /label=A848 tagBFP con -f
                     /note="color: black; sequence: caaggaggccaacaacgaga; added:
                     2022-06-21"
     primer_bind     865..885
                     /label=A897 BFP BsaI -f
                     /note="color: black; sequence: acaacgagacGtacgtcgagc; 
                     added: 2022-07-08"
     primer_bind     866..895
                     /label=BFP BsaI Mut -f
                     /note="color: black; sequence: 
                     CAACGAGACgTACGTCGAGCAGCACGAGGT; added: 2022-07-08"
     primer_bind     926..978
                     /label=BFP-LS-vec-f
                     /note="color: black; sequence: 
                     CAAACTGGGGCACAAGCTTAATTAAtaagctagaactatagtgagtcgtatta; 
                     added: 2022-08-16"
     primer_bind     complement(926..970)
                     /label=BFP-LS-ins-r
                     /note="color: black; sequence: 
                     ctcactatagttctagcttaTTAATTAAGCTTGTGCCCCAGTTTG; added: 
                     2022-08-16"
     primer_bind     complement(926..950)
                     /label=C248 BFP NotI -r
                     /note="color: black; sequence: 
                     aggcggccgcttaattaagcttgtgccccagtttg; added: 2022-06-21"
     promoter        complement(961..978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    983..1117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(1161..1194)
                     /label=SV40-pA-BsaI-CTCA-r
                     /note="color: black; sequence: 
                     gatggtctctTGAGgaattaaaaaacctcccacacct; added: 2022-08-02"
     CDS             1554..2345
                     /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      2438..3026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     2927..2946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     3180..3197
                     /label=A757 L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     terminator      3356..3383
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      181 ccggtctcta gtccgctaca aacttctctc tgctgaagca ggctggagac gtggaggaga
      241 accctggacc tagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
      301 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
      361 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
      421 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
      481 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
      541 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
      601 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
      661 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
      721 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat
      781 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
      841 gactggaaag aatcaaggag gccaacaacg agacgtacgt cgagcagcac gaggtggcag
      901 tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattaa taagctagaa
      961 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     1021 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1081 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1141 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcctcaaga gacctctaga
     1201 ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac
     1261 cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt
     1321 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
     1381 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta
     1441 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     1501 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg
     1561 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     1621 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     1681 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     1741 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     1801 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     1861 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     1921 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     1981 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     2041 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     2101 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     2161 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     2221 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     2281 cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca
     2341 aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     2401 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2581 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2821 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3061 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3121 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3181 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3241 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3301 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     3361 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     3421 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcgg
//
