LOCUS       Exported                5364 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000506 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5364)
  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..5364
                     /organism="BBF10K_000506 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..870
                     /codon_start=1
                     /product="killer cell immunoglobulin like receptor, two Ig 
                     domains and long cytoplasmic tail 4 isoform B"
                     /label=KIR2DL4IB
                     /translation="MSMSPTVIILACLGFFLDQSVWAHVGGQDKPFCSAWPSAVVPQGG
                     HVTLRCHYRRGFNIFTLYKKDGVPVPELYNRIFWNSFLISPVTPAHAGTYRCRGFHPHS
                     PTEWSAPSNPLVIMVTGLYEKPSLTARPGPTVRAGENVTLSCSSQSSFDIYHLSREGEA
                     HELRLPAVPSINGTFQADFPLGPATHGETYRCFGSFHGSPYEWSDPSDPLPVSVTGNPS
                     SSWPSPTEPSFKTGIARHLHAVIRYSVAIILFTILPFFLLHRWCSKKKMLL"
     primer_bind     complement(871..895)
                     /label=Gibson-rev
     terminator      1014..1061
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1066..1082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1241..1491
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1494..2249)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2340..2559
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2979..3000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3015..3045
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3053..3069
                     /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     3077..3093
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3166..4011
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4119..4270
                     /label=nnpp promoters
     CDS             4310..5125
                     /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(5235..5278)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5296..5312
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5313..5344
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgagcatg
       61 agccccaccg tgatcatcct ggcctgcctg ggcttcttcc tggaccagag cgtgtgggcc
      121 cacgtgggcg gccaggacaa gcccttctgc agcgcctggc ccagcgccgt ggtgccccag
      181 ggcggccacg tgaccctgag atgccactac agaagaggct tcaacatctt caccctgtac
      241 aagaaggacg gcgtgcccgt gcccgagctg tacaacagaa tcttctggaa cagcttcctg
      301 atcagccccg tgacccccgc ccacgccggc acctacagat gcagaggctt ccacccccac
      361 agccccaccg agtggagcgc ccccagcaac cccctggtga tcatggtgac cggcctgtac
      421 gagaagccca gcctgaccgc cagacccggc cccaccgtga gagccggcga gaacgtgacc
      481 ctgagctgca gcagccagag cagcttcgac atctaccacc tgagcagaga gggcgaggcc
      541 cacgagctga gactgcccgc cgtgcccagc atcaacggca ccttccaggc cgacttcccc
      601 ctgggccccg ccacccacgg cgagacctac agatgcttcg gcagcttcca cggcagcccc
      661 tacgagtgga gcgaccccag cgaccccctg cccgtgagcg tgaccggcaa ccccagcagc
      721 agctggccca gccccaccga gcccagcttc aagaccggca tcgccagaca cctgcacgcc
      781 gtgatcagat acagcgtggc catcatcctg ttcaccatcc tgcccttctt cctgctgcac
      841 agatggtgca gcaagaagaa gatgctgctg ggatccgcca ccaacttcag cctgctactg
      901 gacccaataa aaggccacct attaaatcgc gtatagatag ggataacagg gtaatgtcgc
      961 ttgtccactc ttctaacgct gaggtgtcaa tcgtcggagc cgctgagcaa taactagcat
     1021 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gcatggtcat agctgtttcc
     1081 tgagagcttg gcaggtgtcg cgtatgcatc gatcacgtgt ccacgttctt taatagtgga
     1141 ctcttgttcc aaactggaac aacactcgga tcgatccggc gcgcaccgtg ggaaaaactc
     1201 caggtagagg tacacacgcg gatagccaat tcagagtaat aaactgtgat aatcaaccct
     1261 catcaatgat gacgaactaa cccccgatat caggtcacat gacgaaggga aagagaagga
     1321 aatcaactgt gacaaactgc cctcaaattt ggcttcctta aaaattacag ttcaaaaagt
     1381 atgagaaaat ccatgcaggc tgaaggaaac agcaaaactg tgacaaatta ccctcagtag
     1441 gtcagaacaa atgtgacgaa ccaccctcaa atctgtgaca gataaccctc agactatcct
     1501 gtcgtcatgg aagtgatatc gcggaaggaa aatacgatat gagtcgtctg gcggcctttc
     1561 tttttctcaa tgtatgagag gcgcattgga gttctgctgt tgatctcatt aacacagact
     1621 tgcaggaagc ggcggcggaa gtcaggcata cgctggtaac tttgaggcag ctggtaacgc
     1681 tctatgatcc agtcgatttt cagagacacg atgcctgagc catccggctt acgatactga
     1741 cacagggatt cgtataaacg catggcatac ggattggtga tttcttttgt ttcactaagc
     1801 cgaaactgcg taaaccggtt ctgtaacccg ataaagaagg gaatgagata tgggttgata
     1861 tgtacactgt aaagccctct ggatggactg tgcgcacgtt tgataaacca aggaaaagat
     1921 tcatagcctt tttcatcacc ggcatcctct tcagggcgat aaaaaaccac ttccttcccc
     1981 gcgaaactct tcaatgcctg ccgtatatcc ttactggctt ccgcagaggt caatccgaat
     2041 atttcagcat atttagcaac atggatctcg cagataccgt catgttcctg tagggtgcca
     2101 tcagattttc tgatctggtc aacgaacaga tacagcatac gtttttgatc ccgggagaga
     2161 ctatatgccg cctcagtgag gtcgtttgac tggacgattc gcgggctatt tttacgtttc
     2221 ttgtgattga taaccgctgt ttccgccatg acagatccat gtgaagtgtg acaagttttt
     2281 agattgtcac actaaataaa aaagagtcaa taagcaggga taactttgtg aaaaaacagc
     2341 ttcttctgag ggcaatttgt cacagggtta agggcaattt gtcacagaca ggactgtcat
     2401 ttgagggtga tttgtcacac tgaaagggca atttgtcaca acaccttctg tagaaccagc
     2461 atggataaag gcctacaagg cgctctaaaa aagaagatct aaaaactata aaaaaaataa
     2521 ttataaaaat atccccgtgg ataagtggat aaccccaagg gaagtttttt caggcatcgt
     2581 gtgtaagcag aatatataag tgctgttccc tggtgcttcc tcgctcactc gaaattcccg
     2641 gggatagctt tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac
     2701 ctctagggtc cccaattaat tagtaatata atctattaaa ggtcattcaa aaggtcatcc
     2761 accggatcaa ttcccctgct cgcgcaggct gggtgccaag ctctcgggta acatcaaggc
     2821 ccgatccttg gagcccttgc cctcccgcac gatgatcgtg ccgtgatcga aatccagatc
     2881 cttgacccgc agttgcaaac cctcactgat ccgattcatt aatgcagctg gcacgacagg
     2941 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     3001 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     3061 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cggtacctgt
     3121 cgcggcaacg cgctaacaga cgtagtaaga accaccagca ttgtaatgct ggctaaagtc
     3181 actttcctga gctgtataac gatgagcgat tttacttttt ctggctatga attggcctgc
     3241 tttgtaacac actccggtct atcccgtagc gccgggcata tcctgtcgca atgtgcaaat
     3301 ctcgcggcaa caaccagtga atacttcatt cacaagcctc accgcctgat cgcggcagaa
     3361 actggttata gccaatcaac cgtcgttcgt gcattccgtg aagctgtaaa caaaggtatc
     3421 ctgtctgtag agattgttat cggcgatcac cgtgaacgtc gcgctaacct gtaccggttt
     3481 acaccatcct ttttggcctt cgcacaacaa gccaaaaatg cgctgataga aagcaaatta
     3541 aagatctctt cagcggcaac caaggttaaa gctgttctcg ctaagacatt ggctttattt
     3601 aattttttat ccacaccccc atgtcaaaat gataccccct ccccctgtca ggatgacgtg
     3661 gcaataaaga ataagaagtc acaagttaaa aaaacaaaaa gatcagtttc cggcggtgcc
     3721 ggaacaacca gcctcaaaaa attgacttca tggatcgcta aggcaaaagc aaaggctgac
     3781 aatctgcggt tatccaaaaa acgcactcaa aaacatgagt tcaagcagaa agtagaggcg
     3841 gctgcgcgga aatatgctta cctgaagaac aagcgttcgc ctgatattgg cgggatatca
     3901 aacttcgata acctaccgca ttgcatgacg gtaaacgaag ctcttaatgc ggttttagcc
     3961 aaaaataaag ataacgaaca atggggtata ccggcaggat tcagagggta atgaattgct
     4021 ctaattataa ccatgcatac tttcaacacc tctagtttgc catgaggcaa actcataggt
     4081 gtcctggtaa gaggacactg ttgccaaaac tggacgcccc attattgcaa ttaataaaca
     4141 actaacggac aattctacct aacaataagt ggagttgcgg ccggccgcga tccgctagca
     4201 aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     4261 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     4321 tcaacgggaa acgtcttgct cgcgcccgcg attaaattcc aacatggatg ctgatttata
     4381 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     4441 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     4501 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     4561 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     4621 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     4681 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     4741 tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     4801 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     4861 actgttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     4921 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4981 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     5041 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     5101 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcgagc
     5161 tctgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5221 tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg
     5281 cacctgcacc agtcagtaaa acgacggcca gtagtcaaaa gcctccgacc ggaggctttt
     5341 gacttggttc aggtggagtg ggag
//
