LOCUS       Exported                5193 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000510 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  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..5193
                     /organism="BBF10K_000510 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..699
                     /codon_start=1
                     /product="KLRK1 killer cell lectin like receptor K1"
                     /label=NKG2D
                     /translation="MGWIRGRRSRHSWEMSEFHNYNLDLKKSDFSTRWQKQRCPVVKSK
                     CRENASPFFFCCFIAVAMGIRFIIMVTIWSAVFLNSLFNQEVQIPLTESYCGPCPKNWI
                     CYKNNCYQFFDESKNWYESQASCMSQNASLLKVYSKEDQDLLKLVKSYHWMGLVHIPTN
                     GSWQWEDGSILSPNLLTIIEMQKGDCALYASSFKGYIENCSTPNTYICMQRTV"
     primer_bind     complement(700..724)
                     /label=Gibson-rev
     terminator      843..890
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(895..911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1070..1320
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1323..2078)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2169..2388
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2808..2829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2844..2874
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2882..2898
                     /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     2906..2922
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2995..3840
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3948..4099
                     /label=nnpp promoters
     CDS             4139..4954
                     /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(5064..5107)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5125..5141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5142..5173
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgggctgg
       61 atcagaggca gaagaagcag acacagctgg gagatgagcg agttccacaa ctacaacctg
      121 gacctgaaga agagcgactt cagcaccaga tggcagaagc agagatgccc cgtggtgaag
      181 agcaagtgca gagagaacgc cagccccttc ttcttctgct gcttcatcgc cgtggccatg
      241 ggcatcagat tcatcatcat ggtgaccatc tggagcgccg tgttcctgaa cagcctgttc
      301 aaccaggagg tgcagatccc cctgaccgag agctactgcg gcccctgccc caagaactgg
      361 atctgctaca agaacaactg ctaccagttc ttcgacgaga gcaagaactg gtacgagagc
      421 caggccagct gcatgagcca gaacgccagc ctgctgaagg tgtacagcaa ggaggaccag
      481 gacctgctga agctggtgaa gagctaccac tggatgggcc tggtgcacat ccccaccaac
      541 ggcagctggc agtgggagga cggcagcatc ctgagcccca acctgctgac catcatcgag
      601 atgcagaagg gcgactgcgc cctgtacgcc agcagcttca agggctacat cgagaactgc
      661 agcaccccca acacctacat ctgcatgcag agaaccgtgg gatccgccac caacttcagc
      721 ctgctagaca agccttaacc gtaggcctaa gcttacaccg caaatctagg gataacaggg
      781 taatacgaac gggattgcac tttcacgctg aggtgtcaat cgtcggagcc gctgagcaat
      841 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg catggtcata
      901 gctgtttcct gagagcttgg caggtgtcgc gtatgcatcg atcacgtgtc cacgttcttt
      961 aatagtggac tcttgttcca aactggaaca acactcggat cgatccggcg cgcaccgtgg
     1021 gaaaaactcc aggtagaggt acacacgcgg atagccaatt cagagtaata aactgtgata
     1081 atcaaccctc atcaatgatg acgaactaac ccccgatatc aggtcacatg acgaagggaa
     1141 agagaaggaa atcaactgtg acaaactgcc ctcaaatttg gcttccttaa aaattacagt
     1201 tcaaaaagta tgagaaaatc catgcaggct gaaggaaaca gcaaaactgt gacaaattac
     1261 cctcagtagg tcagaacaaa tgtgacgaac caccctcaaa tctgtgacag ataaccctca
     1321 gactatcctg tcgtcatgga agtgatatcg cggaaggaaa atacgatatg agtcgtctgg
     1381 cggcctttct ttttctcaat gtatgagagg cgcattggag ttctgctgtt gatctcatta
     1441 acacagactt gcaggaagcg gcggcggaag tcaggcatac gctggtaact ttgaggcagc
     1501 tggtaacgct ctatgatcca gtcgattttc agagacacga tgcctgagcc atccggctta
     1561 cgatactgac acagggattc gtataaacgc atggcatacg gattggtgat ttcttttgtt
     1621 tcactaagcc gaaactgcgt aaaccggttc tgtaacccga taaagaaggg aatgagatat
     1681 gggttgatat gtacactgta aagccctctg gatggactgt gcgcacgttt gataaaccaa
     1741 ggaaaagatt catagccttt ttcatcaccg gcatcctctt cagggcgata aaaaaccact
     1801 tccttccccg cgaaactctt caatgcctgc cgtatatcct tactggcttc cgcagaggtc
     1861 aatccgaata tttcagcata tttagcaaca tggatctcgc agataccgtc atgttcctgt
     1921 agggtgccat cagattttct gatctggtca acgaacagat acagcatacg tttttgatcc
     1981 cgggagagac tatatgccgc ctcagtgagg tcgtttgact ggacgattcg cgggctattt
     2041 ttacgtttct tgtgattgat aaccgctgtt tccgccatga cagatccatg tgaagtgtga
     2101 caagttttta gattgtcaca ctaaataaaa aagagtcaat aagcagggat aactttgtga
     2161 aaaaacagct tcttctgagg gcaatttgtc acagggttaa gggcaatttg tcacagacag
     2221 gactgtcatt tgagggtgat ttgtcacact gaaagggcaa tttgtcacaa caccttctgt
     2281 agaaccagca tggataaagg cctacaaggc gctctaaaaa agaagatcta aaaactataa
     2341 aaaaaataat tataaaaata tccccgtgga taagtggata accccaaggg aagttttttc
     2401 aggcatcgtg tgtaagcaga atatataagt gctgttccct ggtgcttcct cgctcactcg
     2461 aaattcccgg ggatagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa
     2521 aaaggggacc tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa
     2581 aggtcatcca ccggatcaat tcccctgctc gcgcaggctg ggtgccaagc tctcgggtaa
     2641 catcaaggcc cgatccttgg agcccttgcc ctcccgcacg atgatcgtgc cgtgatcgaa
     2701 atccagatcc ttgacccgca gttgcaaacc ctcactgatc cgattcatta atgcagctgg
     2761 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     2821 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2881 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     2941 ggtacctgtc gcggcaacgc gctaacagac gtagtaagaa ccaccagcat tgtaatgctg
     3001 gctaaagtca ctttcctgag ctgtataacg atgagcgatt ttactttttc tggctatgaa
     3061 ttggcctgct ttgtaacaca ctccggtcta tcccgtagcg ccgggcatat cctgtcgcaa
     3121 tgtgcaaatc tcgcggcaac aaccagtgaa tacttcattc acaagcctca ccgcctgatc
     3181 gcggcagaaa ctggttatag ccaatcaacc gtcgttcgtg cattccgtga agctgtaaac
     3241 aaaggtatcc tgtctgtaga gattgttatc ggcgatcacc gtgaacgtcg cgctaacctg
     3301 taccggttta caccatcctt tttggccttc gcacaacaag ccaaaaatgc gctgatagaa
     3361 agcaaattaa agatctcttc agcggcaacc aaggttaaag ctgttctcgc taagacattg
     3421 gctttattta attttttatc cacaccccca tgtcaaaatg ataccccctc cccctgtcag
     3481 gatgacgtgg caataaagaa taagaagtca caagttaaaa aaacaaaaag atcagtttcc
     3541 ggcggtgccg gaacaaccag cctcaaaaaa ttgacttcat ggatcgctaa ggcaaaagca
     3601 aaggctgaca atctgcggtt atccaaaaaa cgcactcaaa aacatgagtt caagcagaaa
     3661 gtagaggcgg ctgcgcggaa atatgcttac ctgaagaaca agcgttcgcc tgatattggc
     3721 gggatatcaa acttcgataa cctaccgcat tgcatgacgg taaacgaagc tcttaatgcg
     3781 gttttagcca aaaataaaga taacgaacaa tggggtatac cggcaggatt cagagggtaa
     3841 tgaattgctc taattataac catgcatact ttcaacacct ctagtttgcc atgaggcaaa
     3901 ctcataggtg tcctggtaag aggacactgt tgccaaaact ggacgcccca ttattgcaat
     3961 taataaacaa ctaacggaca attctaccta acaataagtg gagttgcggc cggccgcgat
     4021 ccgctagcaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     4081 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     4141 gagccatatt caacgggaaa cgtcttgctc gcgcccgcga ttaaattcca acatggatgc
     4201 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     4261 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     4321 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     4381 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     4441 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     4501 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     4561 taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata acggtttggt
     4621 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     4681 aatgcataaa ctgttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     4741 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     4801 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     4861 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     4921 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     4981 ctggcgagct ctgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5041 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     5101 tgatttctgc acctgcacca gtcagtaaaa cgacggccag tagtcaaaag cctccgaccg
     5161 gaggcttttg acttggttca ggtggagtgg gag
//
