LOCUS       Exported                5031 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000502 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5031)
  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..5031
                     /organism="BBF10K_000502 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..537
                     /codon_start=1
                     /label=IL15P1
                     /translation="MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPK
                     TEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGD
                     ASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS"
     primer_bind     complement(538..562)
                     /label=Gibson-rev
     terminator      681..728
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(733..749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    908..1158
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1161..1916)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2007..2226
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2646..2667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2682..2712
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2720..2736
                     /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     2744..2760
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2833..3678
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3786..3937
                     /label=nnpp promoters
     CDS             3977..4792
                     /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(4902..4945)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     4963..4979
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4980..5011
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgagaatc
       61 agcaagcccc acctgagaag catcagcatc cagtgctacc tgtgcctgct gctgaacagc
      121 cacttcctga ccgaggccgg catccacgtg ttcatcctgg gctgcttcag cgccggcctg
      181 cccaagaccg aggccaactg ggtgaacgtg atcagcgacc tgaagaagat cgaggacctg
      241 atccagagca tgcacatcga cgccaccctg tacaccgaga gcgacgtgca ccccagctgc
      301 aaggtgaccg ccatgaagtg cttcctgctg gagctgcagg tgatcagcct ggagagcggc
      361 gacgccagca tccacgacac cgtggagaac ctgatcatcc tggccaacaa cagcctgagc
      421 agcaacggca acgtgaccga gagcggctgc aaggagtgcg aggagctgga ggagaagaac
      481 atcaaggagt tcctgcagag cttcgtgcac atcgtgcaga tgttcatcaa caccagcgga
      541 tccgccacca acttcagcct gctacgtatg gggaacacta cactcgttgc cctaagcatt
      601 catataggga taacagggta atccagcagc cgaccatgta gatcgctgag gtgtcaatcg
      661 tcggagccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      721 gttttttgca tggtcatagc tgtttcctga gagcttggca ggtgtcgcgt atgcatcgat
      781 cacgtgtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcggatcg
      841 atccggcgcg caccgtggga aaaactccag gtagaggtac acacgcggat agccaattca
      901 gagtaataaa ctgtgataat caaccctcat caatgatgac gaactaaccc ccgatatcag
      961 gtcacatgac gaagggaaag agaaggaaat caactgtgac aaactgccct caaatttggc
     1021 ttccttaaaa attacagttc aaaaagtatg agaaaatcca tgcaggctga aggaaacagc
     1081 aaaactgtga caaattaccc tcagtaggtc agaacaaatg tgacgaacca ccctcaaatc
     1141 tgtgacagat aaccctcaga ctatcctgtc gtcatggaag tgatatcgcg gaaggaaaat
     1201 acgatatgag tcgtctggcg gcctttcttt ttctcaatgt atgagaggcg cattggagtt
     1261 ctgctgttga tctcattaac acagacttgc aggaagcggc ggcggaagtc aggcatacgc
     1321 tggtaacttt gaggcagctg gtaacgctct atgatccagt cgattttcag agacacgatg
     1381 cctgagccat ccggcttacg atactgacac agggattcgt ataaacgcat ggcatacgga
     1441 ttggtgattt cttttgtttc actaagccga aactgcgtaa accggttctg taacccgata
     1501 aagaagggaa tgagatatgg gttgatatgt acactgtaaa gccctctgga tggactgtgc
     1561 gcacgtttga taaaccaagg aaaagattca tagccttttt catcaccggc atcctcttca
     1621 gggcgataaa aaaccacttc cttccccgcg aaactcttca atgcctgccg tatatcctta
     1681 ctggcttccg cagaggtcaa tccgaatatt tcagcatatt tagcaacatg gatctcgcag
     1741 ataccgtcat gttcctgtag ggtgccatca gattttctga tctggtcaac gaacagatac
     1801 agcatacgtt tttgatcccg ggagagacta tatgccgcct cagtgaggtc gtttgactgg
     1861 acgattcgcg ggctattttt acgtttcttg tgattgataa ccgctgtttc cgccatgaca
     1921 gatccatgtg aagtgtgaca agtttttaga ttgtcacact aaataaaaaa gagtcaataa
     1981 gcagggataa ctttgtgaaa aaacagcttc ttctgagggc aatttgtcac agggttaagg
     2041 gcaatttgtc acagacagga ctgtcatttg agggtgattt gtcacactga aagggcaatt
     2101 tgtcacaaca ccttctgtag aaccagcatg gataaaggcc tacaaggcgc tctaaaaaag
     2161 aagatctaaa aactataaaa aaaataatta taaaaatatc cccgtggata agtggataac
     2221 cccaagggaa gttttttcag gcatcgtgtg taagcagaat atataagtgc tgttccctgg
     2281 tgcttcctcg ctcactcgaa attcccgggg atagctttat gcttgtaaac cgttttgtga
     2341 aaaaattttt aaaataaaaa aggggacctc tagggtcccc aattaattag taatataatc
     2401 tattaaaggt cattcaaaag gtcatccacc ggatcaattc ccctgctcgc gcaggctggg
     2461 tgccaagctc tcgggtaaca tcaaggcccg atccttggag cccttgccct cccgcacgat
     2521 gatcgtgccg tgatcgaaat ccagatcctt gacccgcagt tgcaaaccct cactgatccg
     2581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2641 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     2701 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     2761 catgattacg ccaagcgcgg tacctgtcgc ggcaacgcgc taacagacgt agtaagaacc
     2821 accagcattg taatgctggc taaagtcact ttcctgagct gtataacgat gagcgatttt
     2881 actttttctg gctatgaatt ggcctgcttt gtaacacact ccggtctatc ccgtagcgcc
     2941 gggcatatcc tgtcgcaatg tgcaaatctc gcggcaacaa ccagtgaata cttcattcac
     3001 aagcctcacc gcctgatcgc ggcagaaact ggttatagcc aatcaaccgt cgttcgtgca
     3061 ttccgtgaag ctgtaaacaa aggtatcctg tctgtagaga ttgttatcgg cgatcaccgt
     3121 gaacgtcgcg ctaacctgta ccggtttaca ccatcctttt tggccttcgc acaacaagcc
     3181 aaaaatgcgc tgatagaaag caaattaaag atctcttcag cggcaaccaa ggttaaagct
     3241 gttctcgcta agacattggc tttatttaat tttttatcca cacccccatg tcaaaatgat
     3301 accccctccc cctgtcagga tgacgtggca ataaagaata agaagtcaca agttaaaaaa
     3361 acaaaaagat cagtttccgg cggtgccgga acaaccagcc tcaaaaaatt gacttcatgg
     3421 atcgctaagg caaaagcaaa ggctgacaat ctgcggttat ccaaaaaacg cactcaaaaa
     3481 catgagttca agcagaaagt agaggcggct gcgcggaaat atgcttacct gaagaacaag
     3541 cgttcgcctg atattggcgg gatatcaaac ttcgataacc taccgcattg catgacggta
     3601 aacgaagctc ttaatgcggt tttagccaaa aataaagata acgaacaatg gggtataccg
     3661 gcaggattca gagggtaatg aattgctcta attataacca tgcatacttt caacacctct
     3721 agtttgccat gaggcaaact cataggtgtc ctggtaagag gacactgttg ccaaaactgg
     3781 acgccccatt attgcaatta ataaacaact aacggacaat tctacctaac aataagtgga
     3841 gttgcggccg gccgcgatcc gctagcaaag ccacgttgtg tctcaaaatc tctgatgtta
     3901 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3961 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcgc gcccgcgatt
     4021 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     4081 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4141 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4201 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4261 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     4321 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4381 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg
     4441 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4501 tgaacaagtc tggaaagaaa tgcataaact gttgccattc tcaccggatt cagtcgtcac
     4561 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4621 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4681 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4741 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     4801 ggttaattgg ttgtaacact ggcgagctct gccacctgac gtctaagaaa ccattattat
     4861 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc
     4921 cttagctttc gctaaggatg atttctgcac ctgcaccagt cagtaaaacg acggccagta
     4981 gtcaaaagcc tccgaccgga ggcttttgac ttggttcagg tggagtggga g
//
