LOCUS       Exported                6057 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000537 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6057)
  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..6057
                     /organism="BBF10K_000537 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1563
                     /codon_start=1
                     /product="Homolog of Blimp-1 in T cells isoform 1"
                     /label=Hobit-i1
                     /translation="MKEESAAQLGCCHRPMALGGTGGSLSPSLDFQLFRGDQVFSACRP
                     LPDMVDAHGPSCASWLCPLPLAPGRSALLACLQDLDLNLCTPQPAPLGTDLQGLQEDAL
                     SMKHEPPGLQASSTDDKKFTVKYPQNKDKLGKQPERAGEGAPCPAFSSHNSSSPPPLQN
                     RKSPSPLAFCPCPPVNSISKELPFLLHAFYPGYPLLLPPPHLFTYGALPSDQCPHLLML
                     PQDPSYPTMAMPSLLMMVNELGHPSARWETLLPYPGAFQASGQALPSQARNPGAGAAPT
                     DSPGLERGGMASPAKRVPLSSQTGTAALPYPLKKKNGKILYECNICGKSFGQLSNLKVH
                     LRVHSGERPFQCALCQKSFTQLAHLQKHHLVHTGERPHKCSVCHKRFSSSSNLKTHLRL
                     HSGARPFQCSVCRSRFTQHIHLKLHHRLHAPQPCGLVHTQLPLASLACLAQWHQGALDL
                     MAVASEKHMGYDIDEVKVSSTSQGKARAVSLSSAGTPLVMGQDQNN"
     primer_bind     complement(1564..1588)
                     /label=Gibson-rev
     terminator      1707..1754
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1759..1775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1934..2184
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2187..2942)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3033..3252
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3672..3693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3708..3738
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3746..3762
                     /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     3770..3786
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3859..4704
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4812..4963
                     /label=nnpp promoters
     CDS             5003..5818
                     /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(5928..5971)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5989..6005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6006..6037
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgaaggag
       61 gaatcagctg ctcaactcgg gtgttgtcat cggccaatgg cgctcggtgg tacaggtggg
      121 tcactctctc cttcccttga ttttcaactc tttcgcggag atcaagtatt ttctgcatgt
      181 aggcctcttc ctgatatggt tgatgctcat gggccttcat gtgcttcttg gctctgtcct
      241 ttgccactcg ctccaggacg ttctgctctc cttgcatgtt tgcaagatct tgatttgaat
      301 ctctgtactc cgcaaccagc tccgcttggg actgatcttc aagggttgca agaagatgct
      361 ctctccatga aacatgaacc acctggtctt caggcttcct ctacagatga taagaaattc
      421 acagttaaat atcctcaaaa taaagataaa ctcggaaaac aaccagaaag ggctggtgaa
      481 ggagctcctt gtccagcatt ttcaagtcat aattcatcca gtccgccacc attgcaaaat
      541 cgaaaatccc ctagtccact tgcattttgt ccatgtccgc cagtcaattc aatttctaaa
      601 gaattgcctt ttctcttgca tgcattttat ccagggtatc ctttgttgct cccaccaccg
      661 catctcttta cgtatggggc acttccttca gatcaatgtc ctcatctctt gatgttgcct
      721 caagatcctt cttatcctac tatggctatg ccttcactcc tcatgatggt caatgaactc
      781 ggacatcctt cagcaaggtg ggaaacattg ctcccttatc caggagcatt tcaagcatcc
      841 ggtcaagctc ttccaagtca agcacggaat ccgggagcag gggcagctcc aacagattct
      901 ccgggacttg aacgcggcgg gatggcaagt ccagcaaaac gcgttccact ctctagtcaa
      961 acggggacag ctgcacttcc gtatccgctt aagaagaaga atgggaagat cttgtatgaa
     1021 tgtaatattt gtgggaaaag ttttggtcaa cttagtaacc tcaaagttca tttgagggtt
     1081 cattccggag aacgaccttt tcaatgtgca ctctgtcaga aatcctttac tcaattggct
     1141 catctccaaa agcatcatct tgtacatact ggagaaaggc cacataaatg ttctgtatgt
     1201 cataaacgct tttcatcatc ctcaaatctc aaaactcatc tccgcttgca ttcaggtgca
     1261 cgtccttttc aatgttctgt ttgtcgcagt aggtttacac aacatattca tcttaaactc
     1321 catcataggc tccatgctcc acaaccgtgt ggtttggtac atacgcaact cccgctcgcg
     1381 agtctcgcat gtctcgcaca atggcatcaa ggagctcttg atttgatggc ggtagcttcc
     1441 gaaaagcata tggggtatga tattgatgaa gttaaagttt cttctacgtc acaaggaaaa
     1501 gcgcgtgcag tttctttgtc ctctgcggga acaccactcg tcatggggca ggaccagaac
     1561 aacggatccg ccaccaactt cagcctgcta gtggtctgta aaccgtaccg gcggcttacc
     1621 gctggctgag tagggataac agggtaatgt atttaataaa tagagtcgcc gctgaggtgt
     1681 caatcgtcgg agccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1741 tgaggggttt tttgcatggt catagctgtt tcctgagagc ttggcaggtg tcgcgtatgc
     1801 atcgatcacg tgtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1861 ggatcgatcc ggcgcgcacc gtgggaaaaa ctccaggtag aggtacacac gcggatagcc
     1921 aattcagagt aataaactgt gataatcaac cctcatcaat gatgacgaac taacccccga
     1981 tatcaggtca catgacgaag ggaaagagaa ggaaatcaac tgtgacaaac tgccctcaaa
     2041 tttggcttcc ttaaaaatta cagttcaaaa agtatgagaa aatccatgca ggctgaagga
     2101 aacagcaaaa ctgtgacaaa ttaccctcag taggtcagaa caaatgtgac gaaccaccct
     2161 caaatctgtg acagataacc ctcagactat cctgtcgtca tggaagtgat atcgcggaag
     2221 gaaaatacga tatgagtcgt ctggcggcct ttctttttct caatgtatga gaggcgcatt
     2281 ggagttctgc tgttgatctc attaacacag acttgcagga agcggcggcg gaagtcaggc
     2341 atacgctggt aactttgagg cagctggtaa cgctctatga tccagtcgat tttcagagac
     2401 acgatgcctg agccatccgg cttacgatac tgacacaggg attcgtataa acgcatggca
     2461 tacggattgg tgatttcttt tgtttcacta agccgaaact gcgtaaaccg gttctgtaac
     2521 ccgataaaga agggaatgag atatgggttg atatgtacac tgtaaagccc tctggatgga
     2581 ctgtgcgcac gtttgataaa ccaaggaaaa gattcatagc ctttttcatc accggcatcc
     2641 tcttcagggc gataaaaaac cacttccttc cccgcgaaac tcttcaatgc ctgccgtata
     2701 tccttactgg cttccgcaga ggtcaatccg aatatttcag catatttagc aacatggatc
     2761 tcgcagatac cgtcatgttc ctgtagggtg ccatcagatt ttctgatctg gtcaacgaac
     2821 agatacagca tacgtttttg atcccgggag agactatatg ccgcctcagt gaggtcgttt
     2881 gactggacga ttcgcgggct atttttacgt ttcttgtgat tgataaccgc tgtttccgcc
     2941 atgacagatc catgtgaagt gtgacaagtt tttagattgt cacactaaat aaaaaagagt
     3001 caataagcag ggataacttt gtgaaaaaac agcttcttct gagggcaatt tgtcacaggg
     3061 ttaagggcaa tttgtcacag acaggactgt catttgaggg tgatttgtca cactgaaagg
     3121 gcaatttgtc acaacacctt ctgtagaacc agcatggata aaggcctaca aggcgctcta
     3181 aaaaagaaga tctaaaaact ataaaaaaaa taattataaa aatatccccg tggataagtg
     3241 gataacccca agggaagttt tttcaggcat cgtgtgtaag cagaatatat aagtgctgtt
     3301 ccctggtgct tcctcgctca ctcgaaattc ccggggatag ctttatgctt gtaaaccgtt
     3361 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     3421 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     3481 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttggagccct tgccctcccg
     3541 cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca aaccctcact
     3601 gatccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3661 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3721 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     3781 tatgaccatg attacgccaa gcgcggtacc tgtcgcggca acgcgctaac agacgtagta
     3841 agaaccacca gcattgtaat gctggctaaa gtcactttcc tgagctgtat aacgatgagc
     3901 gattttactt tttctggcta tgaattggcc tgctttgtaa cacactccgg tctatcccgt
     3961 agcgccgggc atatcctgtc gcaatgtgca aatctcgcgg caacaaccag tgaatacttc
     4021 attcacaagc ctcaccgcct gatcgcggca gaaactggtt atagccaatc aaccgtcgtt
     4081 cgtgcattcc gtgaagctgt aaacaaaggt atcctgtctg tagagattgt tatcggcgat
     4141 caccgtgaac gtcgcgctaa cctgtaccgg tttacaccat cctttttggc cttcgcacaa
     4201 caagccaaaa atgcgctgat agaaagcaaa ttaaagatct cttcagcggc aaccaaggtt
     4261 aaagctgttc tcgctaagac attggcttta tttaattttt tatccacacc cccatgtcaa
     4321 aatgataccc cctccccctg tcaggatgac gtggcaataa agaataagaa gtcacaagtt
     4381 aaaaaaacaa aaagatcagt ttccggcggt gccggaacaa ccagcctcaa aaaattgact
     4441 tcatggatcg ctaaggcaaa agcaaaggct gacaatctgc ggttatccaa aaaacgcact
     4501 caaaaacatg agttcaagca gaaagtagag gcggctgcgc ggaaatatgc ttacctgaag
     4561 aacaagcgtt cgcctgatat tggcgggata tcaaacttcg ataacctacc gcattgcatg
     4621 acggtaaacg aagctcttaa tgcggtttta gccaaaaata aagataacga acaatggggt
     4681 ataccggcag gattcagagg gtaatgaatt gctctaatta taaccatgca tactttcaac
     4741 acctctagtt tgccatgagg caaactcata ggtgtcctgg taagaggaca ctgttgccaa
     4801 aactggacgc cccattattg caattaataa acaactaacg gacaattcta cctaacaata
     4861 agtggagttg cggccggccg cgatccgcta gcaaagccac gttgtgtctc aaaatctctg
     4921 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     4981 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgcgccc
     5041 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     5101 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     5161 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     5221 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     5281 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     5341 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     5401 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca
     5461 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     5521 gcctgttgaa caagtctgga aagaaatgca taaactgttg ccattctcac cggattcagt
     5581 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     5641 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     5701 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     5761 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     5821 agaattggtt aattggttgt aacactggcg agctctgcca cctgacgtct aagaaaccat
     5881 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     5941 aaaatcctta gctttcgcta aggatgattt ctgcacctgc accagtcagt aaaacgacgg
     6001 ccagtagtca aaagcctccg accggaggct tttgacttgg ttcaggtgga gtgggag
//
