LOCUS       Exported                2810 bp ds-DNA     circular SYN 18-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pJS-BCD-Strep-PCi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2810
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..20
                     /label=Primer one sequencing
     primer_bind     92..111
                     /label=BG28357
     promoter        112..130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    130..214
                     /label=BCD
                     /label=bicistronic design element
     RBS             148..156
                     /label=SD1
     primer_bind     complement(195..226)
                     /label=BG30305
                     /note="Gibson for PCk kinase domain JS-BCD backbone_rev, 
                     D214A D237A mutagenesis"
     primer_bind     complement(195..214)
                     /label=BG23271
     RBS             200..208
                     /label=SD2
     primer_bind     210..224
                     /label=BG28359
     CDS             215..688
                     /codon_start=1
                     /label=Strep-PCi
                     /translation="MWSHPQFEKSAHRIDVSGPLYLDCQQSGDTVQLIHQTTVLVEPAI
                     ELYGTDTSTTLEIRDFEDENGRCRVNVFTIERDGTLTPWERLASGAVSPAVWSIIERGG
                     ERAALLVPVPYAPLGELTADLVDQFAPQDYTPSRVRIVLPPPEDDKPPPHDDQ"
     CDS             218..241
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
     primer_bind     complement(230..247)
                     /label=BG30309
                     /note="Gibson for PCk TPR JS-BCD backbone_rev"
     primer_bind     247..265
                     /label=Primer 1
     primer_bind     complement(671..688)
                     /label=Primer 2
     primer_bind     686..706
                     /label=BG30308
                     /note="Gibson for PCk TPR JS-BCD backbone_fwd, D214A D237A 
                     mutagenesis"
     primer_bind     686..703
                     /label=BG30304
                     /note="Gibson for PCk kinase domain JS-BCD backbone_fwd"
     primer_bind     688..706
                     /label=BG23272
     terminator      719..741
                     /label=Terminator
     primer_bind     complement(749..768)
                     /label=BG28358
     terminator      820..847
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1024..1569)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     terminator      complement(1683..1777)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1808..2602)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(2691..2738)
                     /label=NEOKAN_promoter
ORIGIN
        1 ggcgcctttt tttatggggg tattttcatc ccaatccaca cgtccaacgc acagcaaaca
       61 ccacgtcgac cctatcagct gcgtgctttc tatgagtcgt tgctgcataa ctaatacgac
      121 tcactatagg ggcccaagtt cacttaaaaa ggagatcaac aatgaaagca attttcgtac
      181 tgaaacatct taatcatgca ggggagggtt tctaatgtgg agccacccgc agttcgagaa
      241 gagcgcgcac aggattgacg tttcgggtcc gctgtacctg gattgtcaac agagcgggga
      301 caccgttcag ctgattcacc aaaccaccgt tctggttgaa ccggcaattg aactatacgg
      361 gaccgatacc agcaccaccc tggaaattcg tgactttgaa gatgaaaacg gaagatgtcg
      421 tgttaacgtt tttaccattg aacgtgacgg taccctgacc ccgtgggaac gtctagcaag
      481 cggagcagtt tcgccggcag tttggagcat tattgagcgt ggaggggagc gtgctgctct
      541 gctggttccg gttccgtacg caccgctggg tgaactgacc gcagacctgg ttgatcaatt
      601 tgcaccgcag gattacaccc cgtcgagggt tagaattgtt ctcccgccgc cggaagacga
      661 taaaccgccg ccgcacgatg accaataatg ccgactcagt tgctgcttct actgggcgcc
      721 ccgcttcggc ggggtttttt tgcaagggat cggttgtcga gtaaggatct ccaggcatcc
      781 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
      841 ttctgcgttt ataggatcct aactcgagcc tagggatata ttccgcttcc tcgctcactg
      901 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
      961 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     1021 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     1081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     1141 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     1201 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     1261 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1321 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     1381 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     1441 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     1501 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     1561 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     1621 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt
     1681 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     1741 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga
     1801 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     1861 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     1921 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     1981 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     2041 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     2101 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     2161 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     2221 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     2281 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     2341 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     2401 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     2461 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     2521 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     2581 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcatg
     2641 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     2701 tcccaacctt accagagggc gccccagctg gcaattccga cgtcgaattc aaaagatctt
     2761 aagtaagtaa gagtatacgt atatcggcta ataacgtatt aaggcgcttc
//
