LOCUS       Exported                5571 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000507 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  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..5571
                     /organism="BBF10K_000507 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1077
                     /codon_start=1
                     /product="killer cell immunoglobulin like receptor, two Ig 
                     domains and long cytoplasmic tail 4 isoform C "
                     /label=KIR2DL4IC
                     /translation="MSMSPTVIILACLGFFLDQSVWAHVGGQDKPFCSAWPSAVVPQGG
                     HVTLRCHYRRGFNIFTLYKKDGVPVPELYNRIFWNSFLISPVTPAHAGTYRCRGFHPHS
                     PTEWSAPSNPLVIMVTGLYEKPSLTARPGPTVRAGENVTLSCSSQSSFDIYHLSREGEA
                     HELRLPAVPSINGTFQADFPLGPATHGETYRCFGSFHGSPYEWSDPSDPLPVSVTGNPS
                     SSWPSPTEPSFKTDAAVMNQEPAGHRTVNREDSDEQDPQEVTYAQLDHCIFTQRKITGP
                     SQRSKRPSTDTSVCIELPNAEPRALSPAHEHHSQALMGSSRETTALSQTQLASSNVPAA
                     GI"
     primer_bind     complement(1078..1102)
                     /label=Gibson-rev
     terminator      1221..1268
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1273..1289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1448..1698
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1701..2456)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2547..2766
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3186..3207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3222..3252
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3260..3276
                     /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     3284..3300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3373..4218
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4326..4477
                     /label=nnpp promoters
     CDS             4517..5332
                     /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(5442..5485)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5503..5519
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5520..5551
                     /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 aagaccgacg ccgccgtgat gaaccaggag
      781 cccgccggcc acagaaccgt gaacagagag gacagcgacg agcaggaccc ccaggaggtg
      841 acctacgccc agctggacca ctgcatcttc acccagagaa agatcaccgg ccccagccag
      901 agaagcaaga gacccagcac cgacaccagc gtgtgcatcg agctgcccaa cgccgagccc
      961 agagccctga gccccgccca cgagcaccac agccaggccc tgatgggcag cagcagagag
     1021 accaccgccc tgagccagac ccagctggcc agcagcaacg tgcccgccgc cggcatcgga
     1081 tccgccacca acttcagcct gctacttcga ttggcaagga ctgaagtaag gcgcttaagt
     1141 gtgttaggga taacagggta attattttat cgttttagtg cttcgctgag gtgtcaatcg
     1201 tcggagccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1261 gttttttgca tggtcatagc tgtttcctga gagcttggca ggtgtcgcgt atgcatcgat
     1321 cacgtgtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcggatcg
     1381 atccggcgcg caccgtggga aaaactccag gtagaggtac acacgcggat agccaattca
     1441 gagtaataaa ctgtgataat caaccctcat caatgatgac gaactaaccc ccgatatcag
     1501 gtcacatgac gaagggaaag agaaggaaat caactgtgac aaactgccct caaatttggc
     1561 ttccttaaaa attacagttc aaaaagtatg agaaaatcca tgcaggctga aggaaacagc
     1621 aaaactgtga caaattaccc tcagtaggtc agaacaaatg tgacgaacca ccctcaaatc
     1681 tgtgacagat aaccctcaga ctatcctgtc gtcatggaag tgatatcgcg gaaggaaaat
     1741 acgatatgag tcgtctggcg gcctttcttt ttctcaatgt atgagaggcg cattggagtt
     1801 ctgctgttga tctcattaac acagacttgc aggaagcggc ggcggaagtc aggcatacgc
     1861 tggtaacttt gaggcagctg gtaacgctct atgatccagt cgattttcag agacacgatg
     1921 cctgagccat ccggcttacg atactgacac agggattcgt ataaacgcat ggcatacgga
     1981 ttggtgattt cttttgtttc actaagccga aactgcgtaa accggttctg taacccgata
     2041 aagaagggaa tgagatatgg gttgatatgt acactgtaaa gccctctgga tggactgtgc
     2101 gcacgtttga taaaccaagg aaaagattca tagccttttt catcaccggc atcctcttca
     2161 gggcgataaa aaaccacttc cttccccgcg aaactcttca atgcctgccg tatatcctta
     2221 ctggcttccg cagaggtcaa tccgaatatt tcagcatatt tagcaacatg gatctcgcag
     2281 ataccgtcat gttcctgtag ggtgccatca gattttctga tctggtcaac gaacagatac
     2341 agcatacgtt tttgatcccg ggagagacta tatgccgcct cagtgaggtc gtttgactgg
     2401 acgattcgcg ggctattttt acgtttcttg tgattgataa ccgctgtttc cgccatgaca
     2461 gatccatgtg aagtgtgaca agtttttaga ttgtcacact aaataaaaaa gagtcaataa
     2521 gcagggataa ctttgtgaaa aaacagcttc ttctgagggc aatttgtcac agggttaagg
     2581 gcaatttgtc acagacagga ctgtcatttg agggtgattt gtcacactga aagggcaatt
     2641 tgtcacaaca ccttctgtag aaccagcatg gataaaggcc tacaaggcgc tctaaaaaag
     2701 aagatctaaa aactataaaa aaaataatta taaaaatatc cccgtggata agtggataac
     2761 cccaagggaa gttttttcag gcatcgtgtg taagcagaat atataagtgc tgttccctgg
     2821 tgcttcctcg ctcactcgaa attcccgggg atagctttat gcttgtaaac cgttttgtga
     2881 aaaaattttt aaaataaaaa aggggacctc tagggtcccc aattaattag taatataatc
     2941 tattaaaggt cattcaaaag gtcatccacc ggatcaattc ccctgctcgc gcaggctggg
     3001 tgccaagctc tcgggtaaca tcaaggcccg atccttggag cccttgccct cccgcacgat
     3061 gatcgtgccg tgatcgaaat ccagatcctt gacccgcagt tgcaaaccct cactgatccg
     3121 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3181 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3241 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3301 catgattacg ccaagcgcgg tacctgtcgc ggcaacgcgc taacagacgt agtaagaacc
     3361 accagcattg taatgctggc taaagtcact ttcctgagct gtataacgat gagcgatttt
     3421 actttttctg gctatgaatt ggcctgcttt gtaacacact ccggtctatc ccgtagcgcc
     3481 gggcatatcc tgtcgcaatg tgcaaatctc gcggcaacaa ccagtgaata cttcattcac
     3541 aagcctcacc gcctgatcgc ggcagaaact ggttatagcc aatcaaccgt cgttcgtgca
     3601 ttccgtgaag ctgtaaacaa aggtatcctg tctgtagaga ttgttatcgg cgatcaccgt
     3661 gaacgtcgcg ctaacctgta ccggtttaca ccatcctttt tggccttcgc acaacaagcc
     3721 aaaaatgcgc tgatagaaag caaattaaag atctcttcag cggcaaccaa ggttaaagct
     3781 gttctcgcta agacattggc tttatttaat tttttatcca cacccccatg tcaaaatgat
     3841 accccctccc cctgtcagga tgacgtggca ataaagaata agaagtcaca agttaaaaaa
     3901 acaaaaagat cagtttccgg cggtgccgga acaaccagcc tcaaaaaatt gacttcatgg
     3961 atcgctaagg caaaagcaaa ggctgacaat ctgcggttat ccaaaaaacg cactcaaaaa
     4021 catgagttca agcagaaagt agaggcggct gcgcggaaat atgcttacct gaagaacaag
     4081 cgttcgcctg atattggcgg gatatcaaac ttcgataacc taccgcattg catgacggta
     4141 aacgaagctc ttaatgcggt tttagccaaa aataaagata acgaacaatg gggtataccg
     4201 gcaggattca gagggtaatg aattgctcta attataacca tgcatacttt caacacctct
     4261 agtttgccat gaggcaaact cataggtgtc ctggtaagag gacactgttg ccaaaactgg
     4321 acgccccatt attgcaatta ataaacaact aacggacaat tctacctaac aataagtgga
     4381 gttgcggccg gccgcgatcc gctagcaaag ccacgttgtg tctcaaaatc tctgatgtta
     4441 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     4501 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcgc gcccgcgatt
     4561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     4621 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4801 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     4861 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg
     4981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     5041 tgaacaagtc tggaaagaaa tgcataaact gttgccattc tcaccggatt cagtcgtcac
     5101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     5161 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     5221 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     5281 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     5341 ggttaattgg ttgtaacact ggcgagctct gccacctgac gtctaagaaa ccattattat
     5401 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc
     5461 cttagctttc gctaaggatg atttctgcac ctgcaccagt cagtaaaacg acggccagta
     5521 gtcaaaagcc tccgaccgga ggcttttgac ttggttcagg tggagtggga g
//
