LOCUS       pMK445_(BSD-P2A-        3843 bp DNA     circular SYN 18-FEB-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3843)
  AUTHORS   Molecular Function Laboratory
  TITLE     Direct Submission
  JOURNAL   Exported Feb 2, 2024 from SnapGene 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3843
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    701..706
                     /label=Kozak seq
                     /note="color: #000000"
     CDS             707..1102
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /note="color: #ccffcc"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1115..1171
                     /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: #ff0000
                     Cleavage site after base 1168"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     1167..1197
                     /label=(2999) BglII-BromoTag
                     /note="color: black; sequence: 
                     GACCTagatctGTGAAGGACGTGCCCGACTC; added: 2022-02-18"
     CDS             1178..1564
                     /codon_start=1
                     /label=BromoTag
                     /note="color: #ff9900"
                     /translation="VKDVPDSQQHPAPEKSSKVSEQLKCCSGILKEMFAKKHAAYAWPF
                     YKPVDVEALGAHDYCDIIKHPMDMSTIKSKLEAREYRDAQEFGADVRLMFSNCYKYNPP
                     DHEVVAMARKLQDVFEMRFAKMPDE"
     modified_base   1343..1345
                     /label=L387
                     /mod_base="OTHER"
                     /note="color: #a6acb3"
     primer_bind     complement(1562..1609)
                     /label=(3000) BromoTag-BamHI
                     /note="color: black; sequence: 
                     agtggatccGGCGCCTGCACCAGCGCCGCCACCTCCGCCTCCGCCCTC; added: 
                     2022-02-18"
     CDS             1565..1600
                     /codon_start=1
                     /label=Linker
                     /note="color: #ff99cc"
                     /translation="GGGGGGGAGAGA"
     promoter        complement(1655..1673)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(1694..1710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    1718..1734
                     /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)."
                     /note="color: #31849b"
     promoter        complement(join(1742..1748,1749..1766,1767..1772))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 1742 .. 1748 / #ffffff / -10
                        2: 1749 .. 1766 / #ffffff
                        3: 1767 .. 1772 / #ffffff / -35"
     protein_bind    1787..1808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(2096..2684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(join(2855..3646,3647..3715))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 2855 .. 3646 / #ccffcc
                        2: 3647 .. 3715 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3716..3820)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gccaccatgg ccaagccttt
      721 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
      781 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
      841 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
      901 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
      961 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     1021 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     1081 ctctggttat gtgtgggagg gctcaggaag cggagctact aacttcagcc tgctgaagca
     1141 ggctggagac gtggaggaga accctggacc tagatctgtg aaggacgtgc ccgactctca
     1201 gcagcaccca gcaccagaga agagcagcaa ggtctcggag cagctcaagt gctgcagcgg
     1261 catcctcaag gagatgtttg ccaagaagca cgccgcctac gcctggccct tctacaagcc
     1321 tgtggacgtg gaggcactgg gcgcccacga ctactgtgac atcatcaagc accccatgga
     1381 catgagcaca atcaagtcta aactggaggc ccgtgagtac cgtgatgctc aggagtttgg
     1441 tgctgacgtc cgattgatgt tctccaactg ctataagtac aaccctcctg accatgaggt
     1501 ggtggccatg gcccgcaagc tccaggatgt gttcgaaatg cgctttgcca agatgccgga
     1561 cgagggcgga ggcggaggtg gcggcgctgg tgcaggcgcc ggatccacta gttctagagc
     1621 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     1681 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1741 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1801 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1861 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2521 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2761 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2821 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3121 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3721 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3781 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3841 cac
//
