LOCUS       Exported                7149 bp ds-DNA     circular SYN 20-APR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    p5B3-T
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7149)
  AUTHORS   Mark Politz
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 21, 2017 from SnapGene 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7149
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(1..816)
                     /codon_start=1
                     /note="kanR"
                     /note="kanamycin resistance gene from TN903"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(913..1575)
                     /codon_start=1
                     /note="rep"
                     /note="rep gene from pBBR1, GenBank Accession Number 
                     X66730"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      2258..2301
                     /note="terminator"
     CDS             complement(2349..2972)
                     /note="TetR"
     RBS             complement(2979..2983)
                     /note="RBS"
     misc_feature    2992..2997
                     /note="-35"
     protein_bind    2997..3015
                     /note="tet operator"
     misc_feature    complement(2997..3002)
                     /note="-10"
     misc_feature    3016..3021
                     /note="-10"
     misc_feature    complement(3020..3025)
                     /note="-35"
     protein_bind    3027..3045
                     /note="tet operator"
     RBS             3068..3073
                     /note="RBS"
     misc_feature    3084..3107
                     /note="His 7x Tag"
     gene            3108..3833
                     /note="TEV Protease"
     misc_feature    3762..3764
                     /note="S219V"
                     /note="S219V, this version of TEV does not autocatalyze and
                     inactivate itself and is more efficient than wildtype (ie 
                     with Serine)"
     misc_feature    3816..3830
                     /note="poly-arginine tag"
                     /note="poly-arginine tag, purification by cation-exchange 
                     chromatography"
     terminator      complement(3902..3938)
                     /note="SoxR terminator"
                     /note="SoxR terminator (what is the directionality of this 
                     terminator?)"
     promoter        4011..4045
                     /note="J23102 promoter"
     misc_feature    4011..4016
                     /note="-35"
     misc_feature    4034..4039
                     /note="-10"
     RBS             4072..4077
                     /note="RBS"
     CDS             4086..7067
                     /codon_start=1
                     /note="TALElysA"
                     /translation="MHHHHHHVDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTH
                     AHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELR
                     GPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNGSGENLYFQSGSGLTPEQVV
                     AIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQA
                     HGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQ
                     RLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGG
                     GKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQ
                     VVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLC
                     QAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALET
                     VQRLLPVLCQAHGLTPEQVVAIASNHGGKQALETVQRLLPVLCQAHGLTPEQVVAIASN
                     GGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTP
                     EQVVAIASNHGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPV
                     LCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQAL
                     ETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIA
                     SNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGL
                     TPEQVVAIASHDGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGL
                     PHAPALIKRTNRRIPERTSHRVADHAQVVRVLGFFQCHSHPAQAFDDAMTQFGMSDYKD
                     DDDK"
     misc_feature    4086..4514
                     /note="TALE N-terminus"
     misc_feature    4515..4553
                     /note="TEV site"
                     /note="TEV site (NtL) (CAI = 0.871)"
     misc_feature    4554..4655
                     /note="Repeat 1 (HD)"
     misc_feature    4656..4757
                     /note="Repeat 2 (NG)"
     misc_feature    4758..4859
                     /note="Repeat 3 (NG)"
     misc_feature    4860..4961
                     /note="Repeat 4 (NG)"
     misc_feature    4962..5063
                     /note="Repeat 5 (NG)"
     misc_feature    5064..5165
                     /note="Repeat 6 (NG)"
     misc_feature    5166..5267
                     /note="Repeat 7 (NI)"
     misc_feature    5268..5369
                     /note="Repeat 8 (NG)"
     misc_feature    5370..5471
                     /note="Repeat 9 (NN)"
     misc_feature    5472..5573
                     /note="Repeat 10 (NI)"
     misc_feature    5574..5675
                     /note="Repeat 11 (NG)"
     misc_feature    5676..5777
                     /note="Repeat 12 (NH)"
     misc_feature    5778..5879
                     /note="Repeat 13 (NG)"
     misc_feature    5880..5981
                     /note="Repeat 14 (NN)"
     misc_feature    5982..6083
                     /note="Repeat 15 (NH)"
     misc_feature    6084..6185
                     /note="Repeat 16 (HD)"
     misc_feature    6186..6287
                     /note="Repeat 17 (NN)"
     misc_feature    6288..6389
                     /note="Repeat 18 (NG)"
     misc_feature    6390..6491
                     /note="Repeat 19 (NI)"
     misc_feature    6492..6593
                     /note="Repeat 20 (NI)"
     misc_feature    6594..6695
                     /note="Repeat 21 (NG)"
     misc_feature    6696..6755
                     /note="Repeat 21.5 (0.5-mer) (HD)"
     misc_feature    6756..7067
                     /note="C-terminus"
     misc_feature    7041..7064
                     /note="FLAG tag"
     terminator      complement(7102..7133)
                     /note="TonB terminator"
ORIGIN
        1 tcagaaaaac tcatcgagca tcaagtgaaa ctgcaattta ttcatatcag gattatcaat
       61 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      121 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      181 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      241 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
      301 gccattacgc tcgtcatcaa aatcactcgc accaaccaaa ccgttattca ttcgtgattg
      361 cgcctgagcg agacgaaata cgcgatcgcc gttaaaagga caattacaaa caggaatcga
      421 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
      481 ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc
      541 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
      601 tagcctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      661 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
      721 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
      781 cctcgagcaa gacgtttccc gttgaatatg gctcatttta gcttccttag ctcctgaaaa
      841 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
      901 acctcttacg tactaccggc gcggcagcgt gacccgtgtc ggcggctcca acggctcgcc
      961 atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc gccgttccca
     1021 ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc cgggctggct
     1081 gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca gggtcgggat
     1141 gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac
     1201 ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc
     1261 attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg agatccagcg
     1321 ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt
     1381 ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg ccacgaggtg
     1441 atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg cctcatgcgc
     1501 tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc cgatttctct
     1561 ggactgcgtg gccatgctta tctccatgcg gtagggtgcc gcacggttgc ggcaccatgc
     1621 gcaatcagct gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag
     1681 tcaattacag attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc
     1741 tgttgcgtac cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata
     1801 tctagttctt tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc
     1861 ggctccccct ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc
     1921 ggccttgccc aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg
     1981 gggggcaggc gggcgggctt cgccttcgac tgcccccact cgcataggct tgggtcgttc
     2041 caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac
     2101 ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc cggaggcttt tccccagaga
     2161 aaattaaaaa aattgatggg gcaaggccgc aggccgcgca gttggagccg gtgggtatgt
     2221 ggtcgaaggc tgggtagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     2281 cgactgagcc tttcgtttta tttgatgcct ggctctagta gtgatctaca ctagcactat
     2341 cagtgttatt aagacccact ttcacattta agttgttttt ctaatccgca tatgatcaat
     2401 tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc gatagcttgt
     2461 cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt agcgacttga
     2521 tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct gagtgcatat
     2581 aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt ttcgagagtt
     2641 tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc gcgatgactt
     2701 agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca gctttcccct
     2761 tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa ggcgtcgagc
     2821 aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc tagcttctgg
     2881 gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc taatgcgctg
     2941 ttaatcactt tacttttatc taatctagac atcattaatt cctaattttt gttgacactc
     3001 tatcgttgat agagttattt taccactccc tatcagtgat agagaaaaga attcaaaaga
     3061 tcttttaaga aggagatata catatgcatc atcatcatca tcatcatgga gaaagcttgt
     3121 ttaaggggcc gcgtgattac aacccgatat cgagcaccat ttgtcatttg acgaatgaat
     3181 ctgatgggca cacaacatcg ttgtatggta ttggatttgg tcccttcatc attacaaaca
     3241 agcacttgtt tagaagaaat aatggaacac tgttggtcca atcactacat ggtgtattca
     3301 aggtcaagaa caccacgact ttgcaacaac acctcattga tgggagggac atgataatta
     3361 ttcgcatgcc taaggatttc ccaccatttc ctcaaaagct gaaatttaga gagccacaaa
     3421 gggaagagcg catatgtctt gtgacaacca acttccaaac taagagcatg tctagcatgg
     3481 tgtcagacac tagttgcaca ttcccttcat ctgatggcat attctggaag cattggattc
     3541 aaaccaagga tgggcagtgt ggcagtccat tagtatcaac tagagatggg ttcattgttg
     3601 gtatacactc agcatcgaat ttcaccaaca caaacaatta tttcacaagc gtgccgaaaa
     3661 acttcatgga attgttgaca aatcaggagg cgcagcagtg ggttagtggt tggcgattaa
     3721 atgctgactc agtattgtgg gggggccata aagttttcat ggtgaaacct gaagagcctt
     3781 ttcagccagt taaggaagcg actcaactca tgaatcgtcg tcgccgtcgc taagtccatc
     3841 agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgcggaa
     3901 caaaactaaa gcgccacaag ggcgctttag tttgttttca gtccagttac gctggagtct
     3961 gaggctcgtc ctgaatgata tcaagcttga attcgcggcc gcttctagag ttgacagcta
     4021 gctcagtcct aggtactgtg ctagctacta gatatcgctc aatactgacc aaggaggtat
     4081 ctacaatgca tcaccatcat caccatgttg atctgcgtac cctgggttat agccagcagc
     4141 agcaagaaaa aatcaaaccg aaagttcgta gcaccgttgc acagcatcat gaagcactgg
     4201 ttggtcatgg ttttacccat gcacatattg ttgcactgag ccagcatccg gcagcactgg
     4261 gcaccgttgc agttaaatat caggatatga ttgcagcact gccggaagca acccatgaag
     4321 caattgttgg tgttggtaaa cagtggtcag gtgcacgtgc cctggaagca ctgctgaccg
     4381 ttgccggtga actgcgtggt ccgcctctgc agctggatac cggtcagctg ctgaaaattg
     4441 caaaacgtgg tggtgttacc gcagttgaag cagttcatgc atggcgtaat gcactgaccg
     4501 gtgcaccgct gaatggttct ggcgaaaacc tgtacttcca gtctggctcg ggtctgacac
     4561 cggaacaggt tgttgcaatt gccagccatg atggtggcaa acaggcactg gaaaccgttc
     4621 agcgtctgct gccggttctg tgtcaggcac atggtctgac ccctgaacag gtggtggcca
     4681 ttgcaagcaa tggtggcggt aaacaagccc tggaaacagt gcagcgcctg ttaccggtgc
     4741 tgtgccaggc ccatggctta actccggaac aggtggtagc gatcgcatca aatggcggag
     4801 gtaaacaggc cttagaaacc gtacagcgct tactgccggt gttatgccag gcgcacggcc
     4861 tgacgccaga acaggtagtg gcaatcgcct caaatggcgg tggaaaacag gcgttagaga
     4921 cagtccagcg cctgctgcct gtattatgtc aagcccatgg cctgacccca gagcaagttg
     4981 ttgcgattgc aagtaatggt gggggtaaac aggcacttga gacagttcaa cgtttactgc
     5041 ctgtactgtg ccaagctcac ggtctgactc cggaacaagt cgtcgcgatt gcgagtaacg
     5101 gtggtggcaa acaagcatta gaaacggtgc aacgcctgct gccagttctt tgccaggctc
     5161 acggtttaac ccctgagcag gttgtagcta ttgcgagtaa cattggtggt aagcaggcgt
     5221 tggaaactgt gcaaagactg ctgcccgtgt tgtgccaagc acatggttta accccagaac
     5281 aagtcgtagc aatcgcaagc aatgggggtg gcaagcaagc gcttgaaaca gtacagcgtt
     5341 tattaccggt actttgtcag gcccacggtc ttacaccaga acaagttgtg gccatagcca
     5401 gtaacaatgg cggaaagcag gctctggaaa cggtacaacg tctgttacct gttctgtgtc
     5461 aagcgcacgg attaacacct gaacaagtag ttgccattgc gtcaaatatc ggaggcaagc
     5521 aggccttgga gacagtgcag agattactgc cagtgttgtg tcaggctcat ggccttacac
     5581 ccgagcaggt cgtggcaatt gcatctaatg gtggcggtaa gcaagcttta gagactgttc
     5641 agagactgct tcctgtcctg tgccaggcac acggacttac gcctgagcaa gtggttgcaa
     5701 tcgcctctaa tcatggtggt aagcaagcac tggaaactgt ccaacgctta cttccggtgc
     5761 tttgtcaagc acacggctta acgccagagc aggtcgtcgc catagccagc aacggtggtg
     5821 gtaaacaggc ccttgaaacg gtccaaagac ttctgccggt cctttgccaa gcgcatgggc
     5881 tgacacctga gcaggtagtc gcgattgcct caaataacgg tgggaagcag gcattagaaa
     5941 cagttcaaag attattacca gtcctgtgtc aggcgcatgg gttaacccca gagcaggtag
     6001 ttgcaatagc atccaatcat ggcggaaaac aagcgttgga aacggttcag cggttattgc
     6061 ctgttttgtg ccaggcgcat ggtttgacac ccgagcaagt ggtagccata gcctcacatg
     6121 atgggggtaa acaagctttg gagacagtac aacggctgct tccagtttta tgtcaggccc
     6181 atggattgac gcctgaacaa gttgtcgcta tcgcaagtaa taatggtggt aaacaagcgc
     6241 ttgaaaccgt tcaacgcctt ctgcctgtgc tttgtcaggc acatggactt acgcctgagc
     6301 aagtggttgc aatcgcctct aacggtggtg gtaagcaagc actggaaact gtccaacgct
     6361 tacttccggt gctttgtcaa gcacacggcc tgacaccgga acaggttgtt gcaattgcca
     6421 gcaatatagg tggcaaacag gcactggaaa ccgttcagcg tctgctgccg gttctgtgtc
     6481 aggcacatgg tctgacacct gagcaggtag tcgcgattgc ctcaaacatc ggtgggaagc
     6541 aggcattaga aacagttcaa agattattac cagtcctgtg tcaggcgcat gggctgactc
     6601 cggaacaagt cgtcgcgatt gcgagtaatg gtggtggcaa acaagcatta gaaacggtgc
     6661 aacgcctgct gccagttctt tgccaggctc acggtttaac acccgaacag gttgtcgcga
     6721 tagcttcaca tgatggtggt cgtccggcac tggaaagcat tgttgcacag ctgagccgtc
     6781 cggatccggc actggcagca ctgaccaatg atcatctggt tgcactggca tgtctgggtg
     6841 gtcgccctgc cctggatgca gttaaaaaag gtctgccgca tgcaccggca ctgattaaac
     6901 gtaccaatcg tcgtattccg gaacgtacca gccatcgtgt tgcagatcat gcacaggttg
     6961 ttcgtgttct gggttttttt cagtgtcata gccatccggc acaggcattt gatgatgcaa
     7021 tgacccagtt tggtatgagc gattataaag atgatgatga taaataaggt accctaatca
     7081 tacctgacct ccatagcaga aagtcaaaaa cctccgaccg gaggcttttg acttgagggg
     7141 gatccgccc
//
