LOCUS       Exported                6117 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000538 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  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..6117
                     /organism="BBF10K_000538 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1623
                     /codon_start=1
                     /product="Homolog of Blimp-1 in T cells isoform 2
                     "
                     /label=Hobit-i2
                     /translation="MKEESAAQLGCCHRPMALGGTGGSLSPSLDFQLFRGDQVFSACRP
                     LPDMVDAHGPSCASWLCPLPLAPGRSALLACLQDLDLNLCTPQPAPLGTDLQGLQEDAL
                     SMKHEPPGLQASSTDDKKFTVKYPQNKDKLGKQPERAGEGAPCPAFSSHNSSSPPPLQN
                     RKSPSPLAFCPCPPVNSISKELPFLLHAFYPGYPLLLPPPHLFTYGALPSDQCPHLLML
                     PQDPSYPTMAMPSLLMMVNELGHPSARWETLLPYPGAFQASGQALPSQARNPGAGAAPT
                     DSPGLERGGMASPAKRVPLSSQTGTAALPYPLKKKNGKILYECNICGKSFGQLSNLKVH
                     LRVHSGERPFQCALCQKSFTQLAHLQKHHLVHTGERPHKCSIPWVPGRNHWKSFQAWRE
                     REVCHKRFSSSSNLKTHLRLHSGARPFQCSVCRSRFTQHIHLKLHHRLHAPQPCGLVHT
                     QLPLASLACLAQWHQGALDLMAVASEKHMGYDIDEVKVSSTSQGKARAVSLSSAGTPLV
                     MGQDQNN"
     primer_bind     complement(1624..1648)
                     /label=Gibson-rev
     terminator      1767..1814
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1819..1835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1994..2244
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2247..3002)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3093..3312
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3732..3753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3768..3798
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3806..3822
                     /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     3830..3846
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3919..4764
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4872..5023
                     /label=nnpp promoters
     CDS             5063..5878
                     /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(5988..6031)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     6049..6065
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6066..6097
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgaaggag
       61 gaatccgcag cacaactcgg atgttgtcat cgccctatgg ctctcggcgg gacgggtgga
      121 tcactttctc cgtcactcga ttttcaactt ttccggggag atcaagtatt tagtgcatgt
      181 cgaccgctcc cagatatggt cgatgcacat gggccttctt gtgcttcttg gctctgtccg
      241 ttgccgcttg caccagggcg gtccgcgctc cttgcttgtc tccaagatct tgatttgaat
      301 ttgtgtacgc ctcaacctgc tcctctcggt actgatttgc aagggctcca agaagatgct
      361 ttgtccatga aacatgaacc gccagggttg caagcatcca gtacagatga taagaaattt
      421 acagtcaaat atccacaaaa taaagataaa cttgggaaac aaccagaacg cgcaggtgaa
      481 ggggcgccat gtccagcatt ttcttcccat aattcttcct caccgccacc tcttcaaaat
      541 cgtaaaagtc cgtcccctct cgcattttgt ccatgtccac ctgtcaattc catttctaaa
      601 gaactccctt ttcttctcca tgcattttat ccaggttatc ctttgcttct cccaccgcca
      661 catcttttca cttatggggc tctcccttcc gatcaatgtc cgcatctcct catgcttcct
      721 caagatccat cttatcctac tatggcaatg ccaagtcttc ttatgatggt caatgaactt
      781 ggtcatccat ctgcaaggtg ggaaacattg ctcccttatc caggagcttt tcaagcgtca
      841 gggcaagcgt tgccaagtca agctcgcaat ccaggggctg gagctgctcc gactgattcc
      901 ccagggctcg aacgaggcgg aatggcaagt ccagctaaac gggttcctct ttcttcacaa
      961 acagggactg ctgctctccc atatccattg aagaagaaga acgggaaaat attgtatgaa
     1021 tgtaatattt gtgggaaaag ctttgggcaa ctctctaatt tgaaagtcca tcttcgcgtc
     1081 catagtggag aacgaccatt tcaatgtgcg ttgtgtcaaa agtcctttac acaattggct
     1141 catctccaga aacatcatct cgtacataca ggagaacggc cgcataaatg ttccattccg
     1201 tgggtcccag gacgaaatca ttggaaatcc tttcaagcgt ggcgggaacg agaagtctgt
     1261 cataaacgct ttagtagctc ctcaaatctt aaaactcatt tgcgacttca tagtggagca
     1321 cgcccatttc aatgttctgt ttgtaggtca cggtttactc aacatattca tcttaaactc
     1381 catcataggc tccatgctcc acaaccttgt ggtcttgtcc atacgcaact ccctttggca
     1441 agtctcgcat gtctcgcaca atggcatcaa ggggcgcttg atctcatggc tgtcgcttca
     1501 gaaaagcata tgggatatga tattgatgaa gtaaaagtat catccacatc ccaaggaaaa
     1561 gcacgggctg tttccctttc ttctgcgggg actccattgg tcatgggtca ggaccagaac
     1621 aacggatccg ccaccaactt cagcctgcta gtgttttcca ttttccgcgc taactattcc
     1681 gtttaggtca tagggataac agggtaatta agtaggtatg gcaggcagac gctgaggtgt
     1741 caatcgtcgg agccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1801 tgaggggttt tttgcatggt catagctgtt tcctgagagc ttggcaggtg tcgcgtatgc
     1861 atcgatcacg tgtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1921 ggatcgatcc ggcgcgcacc gtgggaaaaa ctccaggtag aggtacacac gcggatagcc
     1981 aattcagagt aataaactgt gataatcaac cctcatcaat gatgacgaac taacccccga
     2041 tatcaggtca catgacgaag ggaaagagaa ggaaatcaac tgtgacaaac tgccctcaaa
     2101 tttggcttcc ttaaaaatta cagttcaaaa agtatgagaa aatccatgca ggctgaagga
     2161 aacagcaaaa ctgtgacaaa ttaccctcag taggtcagaa caaatgtgac gaaccaccct
     2221 caaatctgtg acagataacc ctcagactat cctgtcgtca tggaagtgat atcgcggaag
     2281 gaaaatacga tatgagtcgt ctggcggcct ttctttttct caatgtatga gaggcgcatt
     2341 ggagttctgc tgttgatctc attaacacag acttgcagga agcggcggcg gaagtcaggc
     2401 atacgctggt aactttgagg cagctggtaa cgctctatga tccagtcgat tttcagagac
     2461 acgatgcctg agccatccgg cttacgatac tgacacaggg attcgtataa acgcatggca
     2521 tacggattgg tgatttcttt tgtttcacta agccgaaact gcgtaaaccg gttctgtaac
     2581 ccgataaaga agggaatgag atatgggttg atatgtacac tgtaaagccc tctggatgga
     2641 ctgtgcgcac gtttgataaa ccaaggaaaa gattcatagc ctttttcatc accggcatcc
     2701 tcttcagggc gataaaaaac cacttccttc cccgcgaaac tcttcaatgc ctgccgtata
     2761 tccttactgg cttccgcaga ggtcaatccg aatatttcag catatttagc aacatggatc
     2821 tcgcagatac cgtcatgttc ctgtagggtg ccatcagatt ttctgatctg gtcaacgaac
     2881 agatacagca tacgtttttg atcccgggag agactatatg ccgcctcagt gaggtcgttt
     2941 gactggacga ttcgcgggct atttttacgt ttcttgtgat tgataaccgc tgtttccgcc
     3001 atgacagatc catgtgaagt gtgacaagtt tttagattgt cacactaaat aaaaaagagt
     3061 caataagcag ggataacttt gtgaaaaaac agcttcttct gagggcaatt tgtcacaggg
     3121 ttaagggcaa tttgtcacag acaggactgt catttgaggg tgatttgtca cactgaaagg
     3181 gcaatttgtc acaacacctt ctgtagaacc agcatggata aaggcctaca aggcgctcta
     3241 aaaaagaaga tctaaaaact ataaaaaaaa taattataaa aatatccccg tggataagtg
     3301 gataacccca agggaagttt tttcaggcat cgtgtgtaag cagaatatat aagtgctgtt
     3361 ccctggtgct tcctcgctca ctcgaaattc ccggggatag ctttatgctt gtaaaccgtt
     3421 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     3481 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     3541 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttggagccct tgccctcccg
     3601 cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca aaccctcact
     3661 gatccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3721 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3781 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     3841 tatgaccatg attacgccaa gcgcggtacc tgtcgcggca acgcgctaac agacgtagta
     3901 agaaccacca gcattgtaat gctggctaaa gtcactttcc tgagctgtat aacgatgagc
     3961 gattttactt tttctggcta tgaattggcc tgctttgtaa cacactccgg tctatcccgt
     4021 agcgccgggc atatcctgtc gcaatgtgca aatctcgcgg caacaaccag tgaatacttc
     4081 attcacaagc ctcaccgcct gatcgcggca gaaactggtt atagccaatc aaccgtcgtt
     4141 cgtgcattcc gtgaagctgt aaacaaaggt atcctgtctg tagagattgt tatcggcgat
     4201 caccgtgaac gtcgcgctaa cctgtaccgg tttacaccat cctttttggc cttcgcacaa
     4261 caagccaaaa atgcgctgat agaaagcaaa ttaaagatct cttcagcggc aaccaaggtt
     4321 aaagctgttc tcgctaagac attggcttta tttaattttt tatccacacc cccatgtcaa
     4381 aatgataccc cctccccctg tcaggatgac gtggcaataa agaataagaa gtcacaagtt
     4441 aaaaaaacaa aaagatcagt ttccggcggt gccggaacaa ccagcctcaa aaaattgact
     4501 tcatggatcg ctaaggcaaa agcaaaggct gacaatctgc ggttatccaa aaaacgcact
     4561 caaaaacatg agttcaagca gaaagtagag gcggctgcgc ggaaatatgc ttacctgaag
     4621 aacaagcgtt cgcctgatat tggcgggata tcaaacttcg ataacctacc gcattgcatg
     4681 acggtaaacg aagctcttaa tgcggtttta gccaaaaata aagataacga acaatggggt
     4741 ataccggcag gattcagagg gtaatgaatt gctctaatta taaccatgca tactttcaac
     4801 acctctagtt tgccatgagg caaactcata ggtgtcctgg taagaggaca ctgttgccaa
     4861 aactggacgc cccattattg caattaataa acaactaacg gacaattcta cctaacaata
     4921 agtggagttg cggccggccg cgatccgcta gcaaagccac gttgtgtctc aaaatctctg
     4981 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     5041 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgcgccc
     5101 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     5161 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     5221 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     5281 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     5341 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     5401 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     5461 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca
     5521 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     5581 gcctgttgaa caagtctgga aagaaatgca taaactgttg ccattctcac cggattcagt
     5641 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     5701 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     5761 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     5821 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     5881 agaattggtt aattggttgt aacactggcg agctctgcca cctgacgtct aagaaaccat
     5941 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     6001 aaaatcctta gctttcgcta aggatgattt ctgcacctgc accagtcagt aaaacgacgg
     6061 ccagtagtca aaagcctccg accggaggct tttgacttgg ttcaggtgga gtgggag
//
