LOCUS       Exported                5340 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000534 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  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..5340
                     /organism="BBF10K_000534 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..846
                     /codon_start=1
                     /product="C-lectin type domain family 12 member A"
                     /label=CLEC12A
                     /translation="MSEEVTYADLQFQNSSEMEKIPEIGKFGEKAPPAPSHVWRPAALF
                     LTLLCLLLLIGLGVLASMFHVTLKIEMKKMNKLQNISEELQRNISLQLMSNMNISNKIR
                     NLSTTLQTIATKLCRELYSKEQEHKCKPCPRRWIWHKDSCYFLSDDVQTWQESKMACAA
                     QNASLLKINNKNALEFIKSQSRSYDYWLGLSPEEDSTRGMRVDNIINSSAWVIRNAPDL
                     NNMYCGYINRLYVQYYHCTYKKRMICEKMANPVQLGSTYFREA"
     primer_bind     complement(847..871)
                     /label=Gibson-rev
     terminator      990..1037
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1042..1058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1217..1467
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1470..2225)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2316..2535
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2955..2976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2991..3021
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3029..3045
                     /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     3053..3069
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3142..3987
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4095..4246
                     /label=nnpp promoters
     CDS             4286..5101
                     /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(5211..5254)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5272..5288
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5289..5320
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgagcgag
       61 gaagttactt atgcagattt gcaatttcaa aattcttccg aaatggagaa aattccggaa
      121 attggaaaat ttggagaaaa ggcaccgcca gcgccatcac atgtatggcg accagctgca
      181 ctctttctta cattgctctg tctcttgctc ctgataggac tcggtgtctt ggcatctatg
      241 tttcatgtca cattgaaaat agaaatgaag aaaatgaata aattgcaaaa catttctgaa
      301 gaactccaac ggaatatttc tctccaactc atgtccaaca tgaatattag taataaaatc
      361 cggaatctct ctactactct tcaaactatt gcaacaaaac tctgtcgaga actctattcc
      421 aaagaacaag aacataaatg taaaccatgt cctcgcaggt ggatatggca taaagactcc
      481 tgttattttc ttagtgatga tgttcaaact tggcaagaaa gtaaaatggc atgtgctgcg
      541 caaaatgcta gtctcttgaa aattaataat aagaacgcgc tcgaatttat taagagtcaa
      601 tcacggtcat atgattattg gctcggtctc tctcctgaag aagatagtac tcgcggtatg
      661 cgggttgata atattattaa ttcttccgct tgggtcattc gaaatgctcc tgaccttaat
      721 aatatgtatt gtgggtatat taatcggctt tatgtccaat attatcactg cacatataag
      781 aaaaggatga tttgtgaaaa gatggctaac ccagtacaat tggggtccac atattttaga
      841 gaggccggat ccgccaccaa cttcagcctg ctagtcccaa gttcagacgt actcaagcgc
      901 aaagaggctc aaatagggat aacagggtaa ttgccaacgt ggcgtttaat tgcgctgagg
      961 tgtcaatcgt cggagccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1021 tcttgagggg ttttttgcat ggtcatagct gtttcctgag agcttggcag gtgtcgcgta
     1081 tgcatcgatc acgtgtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1141 ctcggatcga tccggcgcgc accgtgggaa aaactccagg tagaggtaca cacgcggata
     1201 gccaattcag agtaataaac tgtgataatc aaccctcatc aatgatgacg aactaacccc
     1261 cgatatcagg tcacatgacg aagggaaaga gaaggaaatc aactgtgaca aactgccctc
     1321 aaatttggct tccttaaaaa ttacagttca aaaagtatga gaaaatccat gcaggctgaa
     1381 ggaaacagca aaactgtgac aaattaccct cagtaggtca gaacaaatgt gacgaaccac
     1441 cctcaaatct gtgacagata accctcagac tatcctgtcg tcatggaagt gatatcgcgg
     1501 aaggaaaata cgatatgagt cgtctggcgg cctttctttt tctcaatgta tgagaggcgc
     1561 attggagttc tgctgttgat ctcattaaca cagacttgca ggaagcggcg gcggaagtca
     1621 ggcatacgct ggtaactttg aggcagctgg taacgctcta tgatccagtc gattttcaga
     1681 gacacgatgc ctgagccatc cggcttacga tactgacaca gggattcgta taaacgcatg
     1741 gcatacggat tggtgatttc ttttgtttca ctaagccgaa actgcgtaaa ccggttctgt
     1801 aacccgataa agaagggaat gagatatggg ttgatatgta cactgtaaag ccctctggat
     1861 ggactgtgcg cacgtttgat aaaccaagga aaagattcat agcctttttc atcaccggca
     1921 tcctcttcag ggcgataaaa aaccacttcc ttccccgcga aactcttcaa tgcctgccgt
     1981 atatccttac tggcttccgc agaggtcaat ccgaatattt cagcatattt agcaacatgg
     2041 atctcgcaga taccgtcatg ttcctgtagg gtgccatcag attttctgat ctggtcaacg
     2101 aacagataca gcatacgttt ttgatcccgg gagagactat atgccgcctc agtgaggtcg
     2161 tttgactgga cgattcgcgg gctattttta cgtttcttgt gattgataac cgctgtttcc
     2221 gccatgacag atccatgtga agtgtgacaa gtttttagat tgtcacacta aataaaaaag
     2281 agtcaataag cagggataac tttgtgaaaa aacagcttct tctgagggca atttgtcaca
     2341 gggttaaggg caatttgtca cagacaggac tgtcatttga gggtgatttg tcacactgaa
     2401 agggcaattt gtcacaacac cttctgtaga accagcatgg ataaaggcct acaaggcgct
     2461 ctaaaaaaga agatctaaaa actataaaaa aaataattat aaaaatatcc ccgtggataa
     2521 gtggataacc ccaagggaag ttttttcagg catcgtgtgt aagcagaata tataagtgct
     2581 gttccctggt gcttcctcgc tcactcgaaa ttcccgggga tagctttatg cttgtaaacc
     2641 gttttgtgaa aaaattttta aaataaaaaa ggggacctct agggtcccca attaattagt
     2701 aatataatct attaaaggtc attcaaaagg tcatccaccg gatcaattcc cctgctcgcg
     2761 caggctgggt gccaagctct cgggtaacat caaggcccga tccttggagc ccttgccctc
     2821 ccgcacgatg atcgtgccgt gatcgaaatc cagatccttg acccgcagtt gcaaaccctc
     2881 actgatccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2941 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     3001 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     3061 agctatgacc atgattacgc caagcgcggt acctgtcgcg gcaacgcgct aacagacgta
     3121 gtaagaacca ccagcattgt aatgctggct aaagtcactt tcctgagctg tataacgatg
     3181 agcgatttta ctttttctgg ctatgaattg gcctgctttg taacacactc cggtctatcc
     3241 cgtagcgccg ggcatatcct gtcgcaatgt gcaaatctcg cggcaacaac cagtgaatac
     3301 ttcattcaca agcctcaccg cctgatcgcg gcagaaactg gttatagcca atcaaccgtc
     3361 gttcgtgcat tccgtgaagc tgtaaacaaa ggtatcctgt ctgtagagat tgttatcggc
     3421 gatcaccgtg aacgtcgcgc taacctgtac cggtttacac catccttttt ggccttcgca
     3481 caacaagcca aaaatgcgct gatagaaagc aaattaaaga tctcttcagc ggcaaccaag
     3541 gttaaagctg ttctcgctaa gacattggct ttatttaatt ttttatccac acccccatgt
     3601 caaaatgata ccccctcccc ctgtcaggat gacgtggcaa taaagaataa gaagtcacaa
     3661 gttaaaaaaa caaaaagatc agtttccggc ggtgccggaa caaccagcct caaaaaattg
     3721 acttcatgga tcgctaaggc aaaagcaaag gctgacaatc tgcggttatc caaaaaacgc
     3781 actcaaaaac atgagttcaa gcagaaagta gaggcggctg cgcggaaata tgcttacctg
     3841 aagaacaagc gttcgcctga tattggcggg atatcaaact tcgataacct accgcattgc
     3901 atgacggtaa acgaagctct taatgcggtt ttagccaaaa ataaagataa cgaacaatgg
     3961 ggtataccgg caggattcag agggtaatga attgctctaa ttataaccat gcatactttc
     4021 aacacctcta gtttgccatg aggcaaactc ataggtgtcc tggtaagagg acactgttgc
     4081 caaaactgga cgccccatta ttgcaattaa taaacaacta acggacaatt ctacctaaca
     4141 ataagtggag ttgcggccgg ccgcgatccg ctagcaaagc cacgttgtgt ctcaaaatct
     4201 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     4261 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgcg
     4321 cccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     4381 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     4441 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     4501 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     4561 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
     4621 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     4681 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc
     4741 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     4801 ctggcctgtt gaacaagtct ggaaagaaat gcataaactg ttgccattct caccggattc
     4861 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     4921 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     4981 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     5041 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     5101 atcagaattg gttaattggt tgtaacactg gcgagctctg ccacctgacg tctaagaaac
     5161 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     5221 aaaaaaatcc ttagctttcg ctaaggatga tttctgcacc tgcaccagtc agtaaaacga
     5281 cggccagtag tcaaaagcct ccgaccggag gcttttgact tggttcaggt ggagtgggag
//
