LOCUS       Exported                5676 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000505 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  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..5676
                     /organism="BBF10K_000505 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1182
                     /codon_start=1
                     /product="killer cell immunoglobulin like receptor, two Ig 
                     domains and long cytoplasmic tail 4 isoform A"
                     /label=KIR2DL4IA
                     /translation="MSMSPTVIILACLGFFLDQSVWAHVGGQDKPFCSAWPSAVVPQGG
                     HVTLRCHYRRGFNIFTLYKKDGVPVPELYNRIFWNSFLISPVTPAHAGTYRCRGFHPHS
                     PTEWSAPSNPLVIMVTGLYEKPSLTARPGPTVRAGENVTLSCSSQSSFDIYHLSREGEA
                     HELRLPAVPSINGTFQADFPLGPATHGETYRCFGSFHGSPYEWSDPSDPLPVSVTGNPS
                     SSWPSPTEPSFKTGIARHLHAVIRYSVAIILFTILPFFLLHRWCSKKKNAAVMNQEPAG
                     HRTVNREDSDEQDPQEVTYAQLDHCIFTQRKITGPSQRSKRPSTDTSVCIELPNAEPRA
                     LSPAHEHHSQALMGSSRETTALSQTQLASSNVPAAGI"
     primer_bind     complement(1183..1207)
                     /label=Gibson-rev
     terminator      1326..1373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1378..1394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1553..1803
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1806..2561)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2652..2871
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3291..3312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3327..3357
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3365..3381
                     /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     3389..3405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3478..4323
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4431..4582
                     /label=nnpp promoters
     CDS             4622..5437
                     /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(5547..5590)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5608..5624
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5625..5656
                     /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 gaacgccgcc gtgatgaacc aggagcccgc cggccacaga
      901 accgtgaaca gagaggacag cgacgagcag gacccccagg aggtgaccta cgcccagctg
      961 gaccactgca tcttcaccca gagaaagatc accggcccca gccagagaag caagagaccc
     1021 agcaccgaca ccagcgtgtg catcgagctg cccaacgccg agcccagagc cctgagcccc
     1081 gcccacgagc accacagcca ggccctgatg ggcagcagca gagagaccac cgccctgagc
     1141 cagacccagc tggccagcag caacgtgccc gccgccggca tcggatccgc caccaacttc
     1201 agcctgctac tagattagca aggcacccgg gacctgtgga gagtggctat agggataaca
     1261 gggtaattac ctcacctggc aggatcatcg ctgaggtgtc aatcgtcgga gccgctgagc
     1321 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgcatggtc
     1381 atagctgttt cctgagagct tggcaggtgt cgcgtatgca tcgatcacgt gtccacgttc
     1441 tttaatagtg gactcttgtt ccaaactgga acaacactcg gatcgatccg gcgcgcaccg
     1501 tgggaaaaac tccaggtaga ggtacacacg cggatagcca attcagagta ataaactgtg
     1561 ataatcaacc ctcatcaatg atgacgaact aacccccgat atcaggtcac atgacgaagg
     1621 gaaagagaag gaaatcaact gtgacaaact gccctcaaat ttggcttcct taaaaattac
     1681 agttcaaaaa gtatgagaaa atccatgcag gctgaaggaa acagcaaaac tgtgacaaat
     1741 taccctcagt aggtcagaac aaatgtgacg aaccaccctc aaatctgtga cagataaccc
     1801 tcagactatc ctgtcgtcat ggaagtgata tcgcggaagg aaaatacgat atgagtcgtc
     1861 tggcggcctt tctttttctc aatgtatgag aggcgcattg gagttctgct gttgatctca
     1921 ttaacacaga cttgcaggaa gcggcggcgg aagtcaggca tacgctggta actttgaggc
     1981 agctggtaac gctctatgat ccagtcgatt ttcagagaca cgatgcctga gccatccggc
     2041 ttacgatact gacacaggga ttcgtataaa cgcatggcat acggattggt gatttctttt
     2101 gtttcactaa gccgaaactg cgtaaaccgg ttctgtaacc cgataaagaa gggaatgaga
     2161 tatgggttga tatgtacact gtaaagccct ctggatggac tgtgcgcacg tttgataaac
     2221 caaggaaaag attcatagcc tttttcatca ccggcatcct cttcagggcg ataaaaaacc
     2281 acttccttcc ccgcgaaact cttcaatgcc tgccgtatat ccttactggc ttccgcagag
     2341 gtcaatccga atatttcagc atatttagca acatggatct cgcagatacc gtcatgttcc
     2401 tgtagggtgc catcagattt tctgatctgg tcaacgaaca gatacagcat acgtttttga
     2461 tcccgggaga gactatatgc cgcctcagtg aggtcgtttg actggacgat tcgcgggcta
     2521 tttttacgtt tcttgtgatt gataaccgct gtttccgcca tgacagatcc atgtgaagtg
     2581 tgacaagttt ttagattgtc acactaaata aaaaagagtc aataagcagg gataactttg
     2641 tgaaaaaaca gcttcttctg agggcaattt gtcacagggt taagggcaat ttgtcacaga
     2701 caggactgtc atttgagggt gatttgtcac actgaaaggg caatttgtca caacaccttc
     2761 tgtagaacca gcatggataa aggcctacaa ggcgctctaa aaaagaagat ctaaaaacta
     2821 taaaaaaaat aattataaaa atatccccgt ggataagtgg ataaccccaa gggaagtttt
     2881 ttcaggcatc gtgtgtaagc agaatatata agtgctgttc cctggtgctt cctcgctcac
     2941 tcgaaattcc cggggatagc tttatgcttg taaaccgttt tgtgaaaaaa tttttaaaat
     3001 aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta aaggtcattc
     3061 aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca agctctcggg
     3121 taacatcaag gcccgatcct tggagccctt gccctcccgc acgatgatcg tgccgtgatc
     3181 gaaatccaga tccttgaccc gcagttgcaa accctcactg atccgattca ttaatgcagc
     3241 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3301 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3361 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3421 cgcggtacct gtcgcggcaa cgcgctaaca gacgtagtaa gaaccaccag cattgtaatg
     3481 ctggctaaag tcactttcct gagctgtata acgatgagcg attttacttt ttctggctat
     3541 gaattggcct gctttgtaac acactccggt ctatcccgta gcgccgggca tatcctgtcg
     3601 caatgtgcaa atctcgcggc aacaaccagt gaatacttca ttcacaagcc tcaccgcctg
     3661 atcgcggcag aaactggtta tagccaatca accgtcgttc gtgcattccg tgaagctgta
     3721 aacaaaggta tcctgtctgt agagattgtt atcggcgatc accgtgaacg tcgcgctaac
     3781 ctgtaccggt ttacaccatc ctttttggcc ttcgcacaac aagccaaaaa tgcgctgata
     3841 gaaagcaaat taaagatctc ttcagcggca accaaggtta aagctgttct cgctaagaca
     3901 ttggctttat ttaatttttt atccacaccc ccatgtcaaa atgatacccc ctccccctgt
     3961 caggatgacg tggcaataaa gaataagaag tcacaagtta aaaaaacaaa aagatcagtt
     4021 tccggcggtg ccggaacaac cagcctcaaa aaattgactt catggatcgc taaggcaaaa
     4081 gcaaaggctg acaatctgcg gttatccaaa aaacgcactc aaaaacatga gttcaagcag
     4141 aaagtagagg cggctgcgcg gaaatatgct tacctgaaga acaagcgttc gcctgatatt
     4201 ggcgggatat caaacttcga taacctaccg cattgcatga cggtaaacga agctcttaat
     4261 gcggttttag ccaaaaataa agataacgaa caatggggta taccggcagg attcagaggg
     4321 taatgaattg ctctaattat aaccatgcat actttcaaca cctctagttt gccatgaggc
     4381 aaactcatag gtgtcctggt aagaggacac tgttgccaaa actggacgcc ccattattgc
     4441 aattaataaa caactaacgg acaattctac ctaacaataa gtggagttgc ggccggccgc
     4501 gatccgctag caaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa
     4561 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     4621 tatgagccat attcaacggg aaacgtcttg ctcgcgcccg cgattaaatt ccaacatgga
     4681 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     4741 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     4801 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     4861 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     4921 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     4981 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     5041 ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt
     5101 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     5161 agaaatgcat aaactgttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     5221 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     5281 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     5341 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     5401 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     5461 acactggcga gctctgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5521 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
     5581 ggatgatttc tgcacctgca ccagtcagta aaacgacggc cagtagtcaa aagcctccga
     5641 ccggaggctt ttgacttggt tcaggtggag tgggag
//
