LOCUS       Exported                4950 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000503 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
COMMENT     This file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     source          1..4950
                     /organism="BBF10K_000503 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..456
                     /codon_start=1
                     /product="Interleukin 15 Precursor 2"
                     /label=IL15P2
                     /note="Interleukin 15 Precursor 2"
                     /translation="MVLGTIDLCSCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDAT
                     LYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTES
                     GCKECEELEEKNIKEFLQSFVHIVQMFINTS"
     primer_bind     complement(457..481)
                     /label=Gibson-rev
     terminator      600..647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(652..668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    827..1077
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1080..1835)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      1926..2145
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2565..2586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2601..2631
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2639..2655
                     /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     2663..2679
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2752..3597
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3705..3856
                     /label=nnpp promoters
     CDS             3896..4711
                     /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(4821..4864)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     4882..4898
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4899..4930
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catggtgctg
       61 ggcaccatcg acctgtgcag ctgcttcagc gccggcctgc ccaagaccga ggccaactgg
      121 gtgaacgtga tcagcgacct gaagaagatc gaggacctga tccagagcat gcacatcgac
      181 gccaccctgt acaccgagag cgacgtgcac cccagctgca aggtgaccgc catgaagtgc
      241 ttcctgctgg agctgcaggt gatcagcctg gagagcggcg acgccagcat ccacgacacc
      301 gtggagaacc tgatcatcct ggccaacaac agcctgagca gcaacggcaa cgtgaccgag
      361 agcggctgca aggagtgcga ggagctggag gagaagaaca tcaaggagtt cctgcagagc
      421 ttcgtgcaca tcgtgcagat gttcatcaac accagcggat ccgccaccaa cttcagcctg
      481 ctacgtgaaa ctgtatcgag ccgattactc agcatccaag cagtagggat aacagggtaa
      541 tcgggaccct tggtcccttg tacgctgagg tgtcaatcgt cggagccgct gagcaataac
      601 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcat ggtcatagct
      661 gtttcctgag agcttggcag gtgtcgcgta tgcatcgatc acgtgtccac gttctttaat
      721 agtggactct tgttccaaac tggaacaaca ctcggatcga tccggcgcgc accgtgggaa
      781 aaactccagg tagaggtaca cacgcggata gccaattcag agtaataaac tgtgataatc
      841 aaccctcatc aatgatgacg aactaacccc cgatatcagg tcacatgacg aagggaaaga
      901 gaaggaaatc aactgtgaca aactgccctc aaatttggct tccttaaaaa ttacagttca
      961 aaaagtatga gaaaatccat gcaggctgaa ggaaacagca aaactgtgac aaattaccct
     1021 cagtaggtca gaacaaatgt gacgaaccac cctcaaatct gtgacagata accctcagac
     1081 tatcctgtcg tcatggaagt gatatcgcgg aaggaaaata cgatatgagt cgtctggcgg
     1141 cctttctttt tctcaatgta tgagaggcgc attggagttc tgctgttgat ctcattaaca
     1201 cagacttgca ggaagcggcg gcggaagtca ggcatacgct ggtaactttg aggcagctgg
     1261 taacgctcta tgatccagtc gattttcaga gacacgatgc ctgagccatc cggcttacga
     1321 tactgacaca gggattcgta taaacgcatg gcatacggat tggtgatttc ttttgtttca
     1381 ctaagccgaa actgcgtaaa ccggttctgt aacccgataa agaagggaat gagatatggg
     1441 ttgatatgta cactgtaaag ccctctggat ggactgtgcg cacgtttgat aaaccaagga
     1501 aaagattcat agcctttttc atcaccggca tcctcttcag ggcgataaaa aaccacttcc
     1561 ttccccgcga aactcttcaa tgcctgccgt atatccttac tggcttccgc agaggtcaat
     1621 ccgaatattt cagcatattt agcaacatgg atctcgcaga taccgtcatg ttcctgtagg
     1681 gtgccatcag attttctgat ctggtcaacg aacagataca gcatacgttt ttgatcccgg
     1741 gagagactat atgccgcctc agtgaggtcg tttgactgga cgattcgcgg gctattttta
     1801 cgtttcttgt gattgataac cgctgtttcc gccatgacag atccatgtga agtgtgacaa
     1861 gtttttagat tgtcacacta aataaaaaag agtcaataag cagggataac tttgtgaaaa
     1921 aacagcttct tctgagggca atttgtcaca gggttaaggg caatttgtca cagacaggac
     1981 tgtcatttga gggtgatttg tcacactgaa agggcaattt gtcacaacac cttctgtaga
     2041 accagcatgg ataaaggcct acaaggcgct ctaaaaaaga agatctaaaa actataaaaa
     2101 aaataattat aaaaatatcc ccgtggataa gtggataacc ccaagggaag ttttttcagg
     2161 catcgtgtgt aagcagaata tataagtgct gttccctggt gcttcctcgc tcactcgaaa
     2221 ttcccgggga tagctttatg cttgtaaacc gttttgtgaa aaaattttta aaataaaaaa
     2281 ggggacctct agggtcccca attaattagt aatataatct attaaaggtc attcaaaagg
     2341 tcatccaccg gatcaattcc cctgctcgcg caggctgggt gccaagctct cgggtaacat
     2401 caaggcccga tccttggagc ccttgccctc ccgcacgatg atcgtgccgt gatcgaaatc
     2461 cagatccttg acccgcagtt gcaaaccctc actgatccga ttcattaatg cagctggcac
     2521 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2581 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2641 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcggt
     2701 acctgtcgcg gcaacgcgct aacagacgta gtaagaacca ccagcattgt aatgctggct
     2761 aaagtcactt tcctgagctg tataacgatg agcgatttta ctttttctgg ctatgaattg
     2821 gcctgctttg taacacactc cggtctatcc cgtagcgccg ggcatatcct gtcgcaatgt
     2881 gcaaatctcg cggcaacaac cagtgaatac ttcattcaca agcctcaccg cctgatcgcg
     2941 gcagaaactg gttatagcca atcaaccgtc gttcgtgcat tccgtgaagc tgtaaacaaa
     3001 ggtatcctgt ctgtagagat tgttatcggc gatcaccgtg aacgtcgcgc taacctgtac
     3061 cggtttacac catccttttt ggccttcgca caacaagcca aaaatgcgct gatagaaagc
     3121 aaattaaaga tctcttcagc ggcaaccaag gttaaagctg ttctcgctaa gacattggct
     3181 ttatttaatt ttttatccac acccccatgt caaaatgata ccccctcccc ctgtcaggat
     3241 gacgtggcaa taaagaataa gaagtcacaa gttaaaaaaa caaaaagatc agtttccggc
     3301 ggtgccggaa caaccagcct caaaaaattg acttcatgga tcgctaaggc aaaagcaaag
     3361 gctgacaatc tgcggttatc caaaaaacgc actcaaaaac atgagttcaa gcagaaagta
     3421 gaggcggctg cgcggaaata tgcttacctg aagaacaagc gttcgcctga tattggcggg
     3481 atatcaaact tcgataacct accgcattgc atgacggtaa acgaagctct taatgcggtt
     3541 ttagccaaaa ataaagataa cgaacaatgg ggtataccgg caggattcag agggtaatga
     3601 attgctctaa ttataaccat gcatactttc aacacctcta gtttgccatg aggcaaactc
     3661 ataggtgtcc tggtaagagg acactgttgc caaaactgga cgccccatta ttgcaattaa
     3721 taaacaacta acggacaatt ctacctaaca ataagtggag ttgcggccgg ccgcgatccg
     3781 ctagcaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     3841 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     3901 ccatattcaa cgggaaacgt cttgctcgcg cccgcgatta aattccaaca tggatgctga
     3961 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     4021 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     4081 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     4141 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     4201 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     4261 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     4321 cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     4381 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     4441 gcataaactg ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     4501 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     4561 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     4621 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     4681 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     4741 gcgagctctg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4801 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     4861 tttctgcacc tgcaccagtc agtaaaacga cggccagtag tcaaaagcct ccgaccggag
     4921 gcttttgact tggttcaggt ggagtgggag
//
