LOCUS       Exported                5373 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000523 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5373)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     This file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     source          1..5373
                     /organism="BBF10K_000523 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     regulatory      46..55
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             52..879
                     /codon_start=1
                     /product="NCR2 natural cytotoxicity triggering receptor 2"
                     /label=NKp44
                     /translation="MAWRALHPLLLLLLLFPGSQAQSKAQVLQSVAGQTLTVRCQYPPT
                     GSLYEKKGWCKEASALVCIRLVTSSKPRTMAWTSRFTIWDDPDAGFFTVTMTDLREEDS
                     GHYWCRIYRPSDNSVSKSVRFYLVVSPASASTQTSWTPRDLVSSQTQTQSCVPPTAGAR
                     QAPESPSTIPVPSQPQNSTLRPGPAAPIALVPVFCGLLVAKSLVLSALLVWWGDIWWKT
                     MMELRSLDTQKATCHLQQVTDLPWTSVSSPVEREILYHTVARTKISDDDDEHTL"
     primer_bind     complement(880..904)
                     /label=Gibson-rev
     terminator      1023..1070
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1075..1091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1250..1500
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1503..2258)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2349..2568
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2988..3009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3024..3054
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3062..3078
                     /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)."
     primer_bind     3086..3102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3175..4020
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4128..4279
                     /label=nnpp promoters
     CDS             4319..5134
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      complement(5244..5287)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5305..5321
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5322..5353
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catggcctgg
       61 cgtgctttgc atccattgtt gcttttgctt cttctctttc ctggttctca agcacaatcc
      121 aaagctcaag tcctgcaatc agttgctgga caaactctca ctgttcgctg tcaatatccg
      181 ccaacgggta gtctttatga aaagaaagga tggtgtaaag aagctagtgc gttggtctgt
      241 attcgtctcg taacatcttc taaacctcga acaatggcat ggacttcaag gtttacgatt
      301 tgggatgatc ctgatgcagg attctttact gtcactatga cggatctcag ggaagaagat
      361 tcaggacatt attggtgtcg catatatagg ccatctgata attccgtttc taaaagtgtc
      421 cgattttatt tggtagtatc accagcatcc gcttccacgc aaactagttg gacacctcgg
      481 gatcttgtca gttctcaaac tcaaacacaa tcttgtgttc cgcctactgc gggtgcgcga
      541 caagctccag aatctccttc cactatacca gtaccatctc aacctcaaaa ttccacattg
      601 cgcccaggac ctgcagcacc aattgcactc gtacctgtat tttgtggtct cttggttgca
      661 aaatctctcg ttctttctgc tttgctcgta tggtggggtg atatttggtg gaaaactatg
      721 atggaactca ggtcacttga tacacagaaa gctacatgtc atctccaaca agtcacggat
      781 cttccttgga caagtgtctc ctcaccagtt gaaagggaaa ttctctatca tactgtcgct
      841 cgtactaaaa tttccgatga tgatgatgag cacaccctgg gatccgccac caacttcagc
      901 ctgctagcta tgatcccggt gtaacaactg gggaacctcg ttgcattagg gataacaggg
      961 taatgccagc tatggcagtg gggaccgctg aggtgtcaat cgtcggagcc gctgagcaat
     1021 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg catggtcata
     1081 gctgtttcct gagagcttgg caggtgtcgc gtatgcatcg atcacgtgtc cacgttcttt
     1141 aatagtggac tcttgttcca aactggaaca acactcggat cgatccggcg cgcaccgtgg
     1201 gaaaaactcc aggtagaggt acacacgcgg atagccaatt cagagtaata aactgtgata
     1261 atcaaccctc atcaatgatg acgaactaac ccccgatatc aggtcacatg acgaagggaa
     1321 agagaaggaa atcaactgtg acaaactgcc ctcaaatttg gcttccttaa aaattacagt
     1381 tcaaaaagta tgagaaaatc catgcaggct gaaggaaaca gcaaaactgt gacaaattac
     1441 cctcagtagg tcagaacaaa tgtgacgaac caccctcaaa tctgtgacag ataaccctca
     1501 gactatcctg tcgtcatgga agtgatatcg cggaaggaaa atacgatatg agtcgtctgg
     1561 cggcctttct ttttctcaat gtatgagagg cgcattggag ttctgctgtt gatctcatta
     1621 acacagactt gcaggaagcg gcggcggaag tcaggcatac gctggtaact ttgaggcagc
     1681 tggtaacgct ctatgatcca gtcgattttc agagacacga tgcctgagcc atccggctta
     1741 cgatactgac acagggattc gtataaacgc atggcatacg gattggtgat ttcttttgtt
     1801 tcactaagcc gaaactgcgt aaaccggttc tgtaacccga taaagaaggg aatgagatat
     1861 gggttgatat gtacactgta aagccctctg gatggactgt gcgcacgttt gataaaccaa
     1921 ggaaaagatt catagccttt ttcatcaccg gcatcctctt cagggcgata aaaaaccact
     1981 tccttccccg cgaaactctt caatgcctgc cgtatatcct tactggcttc cgcagaggtc
     2041 aatccgaata tttcagcata tttagcaaca tggatctcgc agataccgtc atgttcctgt
     2101 agggtgccat cagattttct gatctggtca acgaacagat acagcatacg tttttgatcc
     2161 cgggagagac tatatgccgc ctcagtgagg tcgtttgact ggacgattcg cgggctattt
     2221 ttacgtttct tgtgattgat aaccgctgtt tccgccatga cagatccatg tgaagtgtga
     2281 caagttttta gattgtcaca ctaaataaaa aagagtcaat aagcagggat aactttgtga
     2341 aaaaacagct tcttctgagg gcaatttgtc acagggttaa gggcaatttg tcacagacag
     2401 gactgtcatt tgagggtgat ttgtcacact gaaagggcaa tttgtcacaa caccttctgt
     2461 agaaccagca tggataaagg cctacaaggc gctctaaaaa agaagatcta aaaactataa
     2521 aaaaaataat tataaaaata tccccgtgga taagtggata accccaaggg aagttttttc
     2581 aggcatcgtg tgtaagcaga atatataagt gctgttccct ggtgcttcct cgctcactcg
     2641 aaattcccgg ggatagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa
     2701 aaaggggacc tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa
     2761 aggtcatcca ccggatcaat tcccctgctc gcgcaggctg ggtgccaagc tctcgggtaa
     2821 catcaaggcc cgatccttgg agcccttgcc ctcccgcacg atgatcgtgc cgtgatcgaa
     2881 atccagatcc ttgacccgca gttgcaaacc ctcactgatc cgattcatta atgcagctgg
     2941 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     3001 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     3061 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     3121 ggtacctgtc gcggcaacgc gctaacagac gtagtaagaa ccaccagcat tgtaatgctg
     3181 gctaaagtca ctttcctgag ctgtataacg atgagcgatt ttactttttc tggctatgaa
     3241 ttggcctgct ttgtaacaca ctccggtcta tcccgtagcg ccgggcatat cctgtcgcaa
     3301 tgtgcaaatc tcgcggcaac aaccagtgaa tacttcattc acaagcctca ccgcctgatc
     3361 gcggcagaaa ctggttatag ccaatcaacc gtcgttcgtg cattccgtga agctgtaaac
     3421 aaaggtatcc tgtctgtaga gattgttatc ggcgatcacc gtgaacgtcg cgctaacctg
     3481 taccggttta caccatcctt tttggccttc gcacaacaag ccaaaaatgc gctgatagaa
     3541 agcaaattaa agatctcttc agcggcaacc aaggttaaag ctgttctcgc taagacattg
     3601 gctttattta attttttatc cacaccccca tgtcaaaatg ataccccctc cccctgtcag
     3661 gatgacgtgg caataaagaa taagaagtca caagttaaaa aaacaaaaag atcagtttcc
     3721 ggcggtgccg gaacaaccag cctcaaaaaa ttgacttcat ggatcgctaa ggcaaaagca
     3781 aaggctgaca atctgcggtt atccaaaaaa cgcactcaaa aacatgagtt caagcagaaa
     3841 gtagaggcgg ctgcgcggaa atatgcttac ctgaagaaca agcgttcgcc tgatattggc
     3901 gggatatcaa acttcgataa cctaccgcat tgcatgacgg taaacgaagc tcttaatgcg
     3961 gttttagcca aaaataaaga taacgaacaa tggggtatac cggcaggatt cagagggtaa
     4021 tgaattgctc taattataac catgcatact ttcaacacct ctagtttgcc atgaggcaaa
     4081 ctcataggtg tcctggtaag aggacactgt tgccaaaact ggacgcccca ttattgcaat
     4141 taataaacaa ctaacggaca attctaccta acaataagtg gagttgcggc cggccgcgat
     4201 ccgctagcaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     4261 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     4321 gagccatatt caacgggaaa cgtcttgctc gcgcccgcga ttaaattcca acatggatgc
     4381 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     4441 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     4501 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     4561 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     4621 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     4681 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     4741 taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata acggtttggt
     4801 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     4861 aatgcataaa ctgttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     4921 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     4981 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     5041 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     5101 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     5161 ctggcgagct ctgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5221 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     5281 tgatttctgc acctgcacca gtcagtaaaa cgacggccag tagtcaaaag cctccgaccg
     5341 gaggcttttg acttggttca ggtggagtgg gag
//
