LOCUS       pMVV094_(RO1600_MSCA_#1 4106 bp ds-DNA     circular     27-JUN-2023
DEFINITION  .
KEYWORDS    "creator:Brian Pfleger Group" "marker:NeoR/KanR"
FEATURES             Location/Qualifiers
     misc_feature    1..6
                     /label="AatII"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_fwdcolor="#ff00ff"
                     /note="note: Restriction site used by the Bgl-brick team to define the BEGINNING of the inducible expression casette, which spans from the repressor up to the terminator of cds."
     misc_feature    7..67
                     /label="L3S2P21"
                     /ApEinfo_revcolor="#000000"
                     /ApEinfo_fwdcolor="#000000"
                     /note="note: Voigt's strongest tested synthetic promoter"
     misc_feature    7..250
                     /label="gBlocks part 1"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    68..87
                     /label="AT-rich random 20-bp"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    88..93
                     /label=-35
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    88..122
                     /label="BBa_J23119"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    111..116
                     /label=-10
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    123..123
                     /label="TSS"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    123..156
                     /label="HP4"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    174..179
                     /label="RBS"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
                     /note="note: RBS"
     misc_feature    188..190
                     /label="START"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    194..223
                     /label="10 HisTag"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
                     /note="note: His Tag"
     misc_feature    224..250
                     /label="Floppy linker"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="note: Linking Histag with the protein, very likely in a manner that minimizes any steric effect of the Histag to the protein, which can potentially affect the conformation thus functionality of the protein. Actually the length of this extended HisTag affects positively the overall insulation purpose of the tag."
     CDS             251..961
                     /label="sfGFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="codon_start: 1 note: sfGFP transl_table: 1 translation: SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     misc_feature    962..964
                     /label="STOP"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    965..973
                     /label="Spacer"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: Spacer as is found in the construct used by Voigt team to test their terminator library."
     terminator      974..1063
                     /label="ECK120029600"
                     /ApEinfo_revcolor="#000000"
                     /ApEinfo_fwdcolor="#000000"
                     /note="note: Voigt's strongest tested natural promoter"
     misc_feature    1064..1069
                     /label="AvrII"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_fwdcolor="#ff00ff"
                     /note="note: Restriction site used by the Bgl-brick team to define the END of the inducible expression casette, which spans from the repressor up to the terminator of cds."
     rep_origin      1070..3038
                     /label="pRO1600_ColE1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="label: pRO1600_ColE1"
     misc_feature    3039..3044
                     /label="SpeI (Pseudo-XbaI)"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_fwdcolor="#ff00ff"
     terminator      3051..3145
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: note: transcription terminator from phage lambda"
     misc_feature    3151..3156
                     /label="SacI"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_fwdcolor="#ff00ff"
     CDS             complement(3176..3970)
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="codon_start: 1 gene: aph(3')-II (or nptII) note: confers resistance to neomycin, kanamycin, and G418 (Geneticin®) product: aminoglycoside phosphotransferase from Tn5 transl_table: 1 translation: MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     promoter        complement(4059..4106)
                     /label="NEOKAN_promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
ORIGIN
        1 gacgtcCTCG GTACCAAATT CCAGAAAAGA GGCCTCCCGA AAGGGGGGCC TTTTTTCGTT
       61 TTGGTCCATC TTGAACTAGC ATACATAttg acagctagct cagtcctagg tataatgcta
      121 gcacgtcgac ttatCTcGaG Tgagatattg ttgacgaact cataaagtca agtaggagat
      181 taattccATG ggccatcacc atcaccatca ccatcaccat cacagcagcg gccatatcga
      241 aggtcgtcat AGCAAAGGAG AAGAACTTTT CACTGGAGTT GTCCCAATTC TTGTTGAATT
      301 AGATGGTGAT GTTAATGGGC ACAAATTTTC TGTCCGTGGA GAGGGTGAAG GTGATGCTAC
      361 AAACGGAAAA CTCACCCTTA AATTTATTTG CACTACTGGA AAACTACCTG TTCCGTGGCC
      421 AACACTTGTC ACTACTCTGA CCTATGGTGT TCAATGCTTT TCCCGTTATC CGGATCACAT
      481 GAAACGGCAT GACTTTTTCA AGAGTGCCAT GCCCGAAGGT TATGTACAGG AACGCACTAT
      541 ATCTTTCAAA GATGACGGGA CCTACAAGAC GCGTGCTGAA GTCAAGTTTG AAGGTGATAC
      601 CCTTGTTAAT CGTATCGAGT TAAAGGGTAT TGATTTTAAA GAAGATGGAA ACATTCTTGG
      661 ACACAAACTT GAGTACAACT TTAACTCACA CAATGTATAC ATCACGGCAG ACAAACAAAA
      721 GAATGGAATC AAAGCTAACT TCAAAATTCG CCACAACGTT GAAGATGGTT CCGTTCAACT
      781 AGCAGACCAT TATCAACAAA ATACTCCAAT TGGCGATGGC CCTGTCCTTT TACCAGACAA
      841 CCATTACCTG TCGACACAAT CTGTCCTTTC GAAAGATCCC AACGAAAAGC GTGACCACAT
      901 GGTCCTTCTT GAGTTTGTAA CTGCTGCTGG GATTACACAT ggcatggatg agctctacaa
      961 ataataagaa ttcTTCAGCC AAAAAACTTA AGACCGCCGG TCTTGTCCAC TACCTTGCAG
     1021 TAATGCGGTG GACAGGATCG GCGGTTTTCT TTTCTCTTCT CAAcctaggG ATAATCTCAT
     1081 GACCAAAATC CCTTAACGTG AGTTTTCGTT CCACTGAGCG TCAGACCCCG TAGAAAAGAT
     1141 CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC TGCTGCTTGC AAACAAAAAA
     1201 ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG CTACCAACTC TTTTTCCGAA
     1261 GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTT CTTCTAGTGT AGCCGTAGTT
     1321 AGGCCACCAC TTCAAGAACT CTGTAGCACC GCCTACATAC CTCGCTCTGC TAATCCTGTT
     1381 ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC GGGTTGGACT CAAGACGATA
     1441 GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT TCGTGCACAC AGCCCAGCTT
     1501 GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT GAGCTATGAG AAAGCGCCAC
     1561 GCTTCCCGAA GGGAGAAAGG CGGACAGGCA TCCGGTAAGC GGCAGGGTCG GAACAGGAGA
     1621 GCGCACGAGG GAGCTTCCAG GGGGAAACGC CTGGTATCTT TATAGTCCTG TCGGGTTTCG
     1681 CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA GGGGGGCGGA GCCTATGGAA
     1741 AAACGCCAGC AACGCGGCCG TGAAAGGCAG GCCGGTCCGT GGTGGCCACG GCCTCTAGGC
     1801 CAGATCCAGC GGCATCTGGG TTAGTCGAGC GCGGGCCGCT TCCCATGTCT CACCAGGGCG
     1861 AGCCTGTTTC GCGATCTCAG CATCTGAAAT CTTCCCGGCC TTGCGCTTCG CTGGGGCCTT
     1921 ACCCACCGCC TTGGCGGGCT TCTTCGGTCC AAAACTGAAC AACAGATGTG TGACCTTGCG
     1981 CCCGGTCTTT CGCTGCGCCC ACTCCACCTG TAGCGGGCTG TGCTCGTTGA TCTGCGTCAC
     2041 GGCTGGATCA AGCACTCGCA ACTTGAAGTC CTTGATCGAG GGATACCGGC CTTCCAGTTG
     2101 AAACCACTTT CGCAGCTGGT CAATTTCTAT TTCGCGCTGG CCGATGCTGT CCCATTGCAT
     2161 GAGCAGCTCG TAAAGCCTGA TCGCGTGGGT GCTGTCCATC TTGGCCACGT CAGCCAAGGC
     2221 GTATTTGGTG AACTGTTTGG TGAGTTCCGT CAGGTACGGC AGCATGTCTT TGGTGAACCT
     2281 GAGTTCTACA CGGCCCTCAC CCTCCCGGTA GATGATTGTT TGCACCCAGC CGGTAATCAT
     2341 CACACTCGGT CTTTTCCCCT TGCCATTGGG CTCTTGGGTT AACCGGACTT CCCGCCGTTT
     2401 CAGGCGCAGG GCCGCTTCTT TGAGCTGGTT GTAGGAAGAT TCGATAGGGA CACCCGCCAT
     2461 CGTCGCTATG TCCTCCGCCG TCACTGAATA CATCACTTCA TCGGTGACAG GCTCGCTCCT
     2521 CTTCACCTGG CTAATACAGG CCAGAACGAT CCGCTGTTCC TGAACACTGA GGCGATACGC
     2581 GGCCTCGACC AGGGCATTGC TTTTGTAAAC CATTGGGGGT GAGGCCACGT TCGACATTCC
     2641 TTGTGTATAA GGGGACACTG TATCTGCGTC CCACAATACA ACAAATCCGT CCCTTTACAA
     2701 CAACAAATCC GTCCCTTCTT AACAACAAAT CCGTCCCTTA ATGGCAACAA ATCCGTCCCT
     2761 TTTTAAACTC TACAGGCCAC GGATTACGTG GCCTGTAGAC GTCCTAAAAG GTTTAAAAGG
     2821 GAAAAGGAAG AAAAGGGTGG AAACGCAAAA AACGCACCAC TACGTGGCCC CGTTGGGGCC
     2881 GCATTTGTGC CCCTGAAGGG GCGGGGGAGG CGTCTGGGCA ATCCCCGTTT TACCAGTCCC
     2941 CTATCGCCGC CTGAGAGGGC GCAGGAAGCG AGTAATCAGG GTATCGAGGC GGATTCACCC
     3001 TTGGCGTCCA ACCAGCGGCA CCAGCGGCGC CTGAGAGGac tagtgcttgg attctcacca
     3061 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3121 attactggat ctatcaacag gagtccaagc gagctctcga accccagagt cccgctcaga
     3181 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     3241 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     3301 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     3361 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     3421 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     3481 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     3541 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     3601 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     3661 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     3721 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     3781 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     3841 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     3901 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     3961 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcatgat cccctgcgcc
     4021 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     4081 cagagggcgc cccagctggc aattcc
//