LOCUS       ND4-DdCBE-left_s        7695 bp ds-DNA     circular SYN 01-JUL-2020
DEFINITION  SOD2 MTS_3xHA_left m.11922  TALE_2aa_DddE(149-178)_1xUGI_SOD2 3'UTR
            .
ACCESSION
VERSION     .
KEYWORDS    TALEN-CCR5L18-NtermN1.
SOURCE      synthetic DNA construct
  ORGANISM  human, from Guilinger paper 
            .
REFERENCE   1  (bases 1 to 8058)
  AUTHORS   Guilinger JP, Pattanayak V, Reyon D, Tsai SQ, Sander JD, Joung JK
            and Liu DR
  TITLE     Broad specificity profiling of TALENs results in engineered
            nucleases with improved DNA-cleavage specificity.
  JOURNAL   Nat Methods. 2014 Apr;11(4):429-35. doi: 10.1038/nmeth.2845. Epub
            2014 Feb 16.
  PUBMED    24531420
REFERENCE   2  (bases 1 to 8058)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7695
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /label="pRS-marker"
     primer_bind     198..213
                     /Sequence="GCGCTGCTTCGCGATG"
                     /Hairpin_Tm="55.5"
                     /Tm="59.7"
                     /Self_Dimer_Tm="14.5"
                     /%GC="68.8"
                     /created_by="primer3"
                     /label="BM719"
     primer_bind     209..229
                     /Sequence="CGATGTACGGGCCAGATATAC"
                     /Hairpin_Tm="38.5"
                     /Tm="57.9"
                     /Self_Dimer_Tm="10.9"
                     /%GC="52.4"
                     /created_by="primer3"
                     /label="BM525"
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /label="CMV promoter"
     primer_bind     687..706
                     /Sequence="CCCATTGACGTCAATGGGAG"
                     /Hairpin_Tm="71.5"
                     /Tm="58.6"
                     /Self_Dimer_Tm="59.4"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="BM720"
     primer_bind     769..789
                     /note="Human CMV immediate early promoter, forward primer"
                     /label="CMV-F"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     863..882
                     /note="T7 promoter, forward primer"
                     /label="T7"
     CDS             904..996
                     /created_by="bmok"
                     /label="SOD2-MTS"
     source          <907..>909
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          <907..>909
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /modified_by="hrees"
                     /label="From pHR_087"
     CDS             997..1083
                     /created_by="bmok"
                     /label="3x HA tag "
     source          <997..>1083
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1087..1500
                     /created_by="bmok"
                     /label="N-terminus cannonical "
     CDS             1501..3192
                     /created_by="bmok"
                     /modified_by="bmok"
                     /label="ND4 Left TALE"
     source          <1501..>3192
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             3193..3315
                     /created_by="bmok"
                     /label="C-terminal domain "
     source          <3316..>3321
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(<3316..>3321,<3316..>3321)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             <3322..3411
                     /created_by="bmok"
                     /modified_by="bmok"
                     /label="DddA(1398-1427)"
     source          <3322..>3411
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(<3322..>3411,<3322..>3411)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          <3412..>3672
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(<3412..>3672,<3412..>3672)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          <3412..>3423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          <3412..3423
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="Genscript 2xUGI"
     CDS             3424..3672
                     /created_by="kzhao"
                     /label="UGI"
     source          <3424..>3672
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          <3424..>3672
                     /created_by="jonlevy"
                     /modified_by="jonlevy"
                     /label="Genscript 2xUGI"
     3'UTR           3679..3880
                     /created_by="bmok"
                     /modified_by="bmok"
                     /label="SOD2-3'UTR"
     motif           3855..3859
                     /created_by="bmok"
                     /label="AAUAA motif "
     rep_origin      3926..4354
                     /direction="RIGHT"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     complement(4013..4032)
                     /note="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     4223..4244
                     /note="F1 origin, forward primer"
                     /label="F1ori-F"
     primer_bind     complement(4363..4383)
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /label="pBABE 3'"
     promoter        4368..4698
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     rep_origin      4549..4684
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     4611..4630
                     /note="SV40 promoter/origin, forward primer"
                     /label="SV40pro-F"
     promoter        4746..4793
                     /note="synthetic bacterial promoter"
                     /label="EM7 promoter"
     CDS             4812..5210
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHP
                     GIKAIVKDSDGQPTAVGIRELLPSGYVWEG"
                     /label="BSD"
     polyA_signal    5368..5489
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5405..5424)
                     /note="SV40 polyA, reverse primer"
                     /label="SV40pA-R"
     primer_bind     5459..5478
                     /note="SV40 polyA terminator, reverse primer"
                     /label="EBV-rev"
     primer_bind     complement(5538..5554)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 rev"
     primer_bind     complement(5538..5554)
                     /note="In lacZ gene. Also called M13-rev"
                     /label="M13 Reverse"
     primer_bind     complement(5551..5573)
                     /note="In lacZ gene"
                     /label="M13/pUC Reverse"
     protein_bind    5562..5578
                     /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)."
                     /label="lac operator"
     promoter        complement(5586..5616)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5631..5652
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /label="CAP binding site"
     misc_feature    complement(5663..5679)
                     /Sequence="GAAAGCGGGCAGTGAGC"
                     /Hairpin_Tm="44.7"
                     /Tm="58.7"
                     /Self_Dimer_Tm="None"
                     /%GC="64.7"
                     /created_by="primer3"
                     /label="BM524 "
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     complement(5769..5786)
                     /note="L4440 vector, forward primer"
                     /label="L4440"
     rep_origin      complement(5940..6528)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     complement(6020..6039)
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     CDS             complement(6699..7559)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     7322..7341
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     promoter        complement(7560..7664)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN      
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatggccc tgtcccgtgc ggtttgtggc acctcccgtc aactggctcc ggttctgggt
      961 tatctgggtt cccgtcaaaa acactccctg ccggactacc cgtatgatgt tccggattac
     1021 gctggctacc catacgacgt cccagactac gctggctacc catacgacgt cccagactac
     1081 gctatggaca tcgcggatct gcgtaccctg ggttacagcc agcagcagca ggagaagatc
     1141 aagccgaagg tgcgcagcac cgtggctcag caccacgaag ccctggtggg ccacggtttc
     1201 acccacgctc acatcgtggc cctgagccag cacccagccg cgctgggcac cgtggccgtg
     1261 aaatatcagg acatgattgc tgccctgcca gaggccaccc atgaagctat tgtgggcgtg
     1321 ggcaagcagt ggagcggtgc tcgtgcgctg gaggcgctgc tgaccgtggc tggtgaactg
     1381 cgtggtccgc cgctgcagct ggataccggt cagctgctga aaatcgcgaa acgtggcggt
     1441 gtgaccgcgg tggaagccgt gcatgcttgg cgtaatgctc tgaccggtgc gccgctgaac
     1501 ctgaccccgg aacaggtggt ggcaatcgca agcaataatg gtggtaaaca ggctctggaa
     1561 accgtgcaag ctctgctgcc agttctgtgc caggctcatg gtctgacccc gcagcaggtg
     1621 gtggctattg ccagccatga tggcggcaag caggccctgg agaccgtgca gcgtctgctg
     1681 ccggtgctgt gccaggccca tggcctgacc ccgcagcaag ttgtggctat tgccagcaac
     1741 ggcggtggca aacaggctct ggagaccgtg cagcgcctgt taccggtgct gtgccaagcc
     1801 catggtctga ccccggagca ggtggtggcg atcgctagca acattggtgg caagcaggcc
     1861 ctggaaaccg tgcaggcgct gttgccggtg ctgtgccaag cccatgggct gaccccggaa
     1921 caagttgttg ccattgccag caacaatggt ggcaaacagg ctctggaaac tgtgcaggcc
     1981 ctgcttccgg tgctgtgcca ggcccatgga ttaaccccgg aacaagttgt ggctattgcg
     2041 agcaatggcg gcggcaagca ggcgctggaa accgtgcagg ctctgttacc ggtgctgtgc
     2101 caggcgcatg gcctgacccc ggagcaagtg gtggccatcg ctagcaacat tggcggtaaa
     2161 caggcgctgg agaccgttca gcgtctgtta ccggtgctgt gccaggcaca tggccttacc
     2221 ccggaacaag ttgtggccat tgccagcaac atcggcggca agcaggccct ggaaacggtg
     2281 caggcgctgc tcccggtgct gtgccaggcc catgggttaa ccccggaaca agtggttgct
     2341 attgctagcc atgatggcgg taaacaggcc ctggagaccg ttcagcgcct gctaccggtg
     2401 ctgtgccagg cccatggttt aaccccggaa caggttgttg cgattgctag ccacgatggc
     2461 ggcaagcagg ctctggagac cgttcaggcc ctgctcccgg tgctgtgcca ggcccatggg
     2521 cttaccccgg agcaagttgt tgctattgcc tccaatattg gcggtaaaca ggcgctggaa
     2581 accgttcagg ctctgcttcc ggtgctgtgc caggctcatg gcctcacccc ggaacaagtt
     2641 gtggcgattg cgtcccatga tggcggcaag caggccctgg aaactgtgca ggcgctgcta
     2701 ccggtgctgt gccaggccca tgggctaacc ccggaacagg tggttgcgat tgctagcaac
     2761 aacggcggta aacaggctct ggagactgtt cagcgtctgc ttccggtgct gtgccaggct
     2821 catgggctga ccccgcagca agtggttgct attgccagca atggcggtgg caagcaggcg
     2881 ctggagactg ttcagcgcct gctcccggtg ctgtgccagg ctcatggttt aaccccggag
     2941 caggttgtgg cgatcgccag caatggtggc ggtaaacagg ctctggaaac ggtgcaggcc
     3001 ctgctcccgg tgctgtgcca ggctcatgga ctgaccccgg agcaagttgt tgccattgcg
     3061 tcccacgacg gcggcaagca ggcgctggag acggtgcagg ctctgctccc ggtgctgtgc
     3121 caggctcacg gtctgacccc gcaacaggtg gtggcaatcg caagcaacgg tggtggtcgt
     3181 ccggcactgg agagcattgt ggcgcagctg agccgtccag acccggccct ggcggctctg
     3241 accaacgatc acctggtggc gctggcttgc ctgggcggtc gtccggccct ggatgcggtg
     3301 aagaaaggcc tgggtggatc cgccattcca gtgaagcgcg gcgctaccgg tgaaaccaaa
     3361 gtgtttaccg gtaacagcaa cagcccgaag agcccgacca aaggcggttg ctctggcggc
     3421 tcaactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     3481 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     3541 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     3601 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     3661 atcaagatgc tgtgataaac cacgatcgtt atgctgatca taccctaatg atcccagcaa
     3721 gataatgtcc tgtcttctaa gatgtgcatc aagcctggta catactgaaa accctataag
     3781 gtcctggata atttttgttt gattattcat tgaagaaaca tttattttcc aattgtgtga
     3841 agtttttgac tgttaataaa agaatctgtc aaccatcaaa cttctgaggc ggaaagaacc
     3901 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3961 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4021 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4081 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4141 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4201 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4261 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     4321 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat
     4441 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4621 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4681 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     4741 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     4801 ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     4861 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     4921 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     4981 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     5041 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgt cgacaggtgc
     5101 ttctcgatct gcatcctggg atcaaagcga tagtgaagga cagtgatgga cagccgacgg
     5161 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg
     5221 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     5281 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5341 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5401 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5461 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5521 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5581 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5641 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5701 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5761 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6121 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6361 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6661 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6721 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6781 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6841 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6901 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6961 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7021 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7081 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7141 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7201 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7261 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7321 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7381 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7441 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7501 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7561 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7621 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7681 gtgccacctg acgtc
//
