LOCUS       Exported                6377 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    JMJD1CA-c089
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  TITLE     SGC 2016 plasmids
REFERENCE   2  (bases 1 to 6377)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6377
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             1126..1146
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1147..1164
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1165..1188
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1238..1255
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1308..1326)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1322..1369
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1406..1861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1493..1512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1703..1724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1954..2769)
                     /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"
     primer_bind     2677..2696
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2761..2780
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2891..3479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3380..3399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3633..3650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3665..3807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3733..3752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3893..3915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3909..4100)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4909..5991)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5953..5972
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5992..6069)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6275..6294
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaacct taagaaggag atatactatg ataccacatt cttggatctg tgagaagcat
      121 attttatggc ttaaggatta taagaatagc agtaattgga agcttttcaa agaatgttgg
      181 aaacaaggac agcctgcagt ggtttctggt gtgcataaga aaatgaacat tagcctatgg
      241 aaggcggaat caattagtct tgattttgga gaccaccaag ctgatctcct gaactgcaaa
      301 gatagcatca tttcaaatgc caatgttaag gaattctggg atggttttga agaagtttca
      361 aaacggcaga aaaacaagag tggagaaaca gttgttttaa aattgaaaga ctggccttca
      421 ggagaagact tcaagactat gatgccagca agatacgaag atcttttaaa aagtctgcca
      481 ttgccagaat attgtaatcc agaaggaaaa ttcaatttgg cctctcattt gccaggattt
      541 tttgtacgtc ctgatctagg acccaggttg tgcagtgcct atggtgtagt tgctgctaaa
      601 gatcatgata taggaacaac aaatctccat attgaagttt ctgatgttgt aaatatacta
      661 gtttatgttg gcatagcaaa aggaaatggc attctctcaa aagcaggaat tctcaagaaa
      721 tttgaggaag aagatttgga tgacatttta aggaaaagat tgaaggactc aagtgaaata
      781 cctggtgctc tgtggcatat ttatgctggg aaagatgttg acaagataag ggaatttctt
      841 caaaagattt caaaagaaca aggccttgaa gttctaccag aacatgatcc aatacgtgac
      901 caaagttggt atgtgaacaa aaagctccgt caaaggctgc ttgaagaata tggagtcaga
      961 acctgtactc ttattcagtt ccttggtgat gctattgttt tgccagcggg agcacttcat
     1021 caggttcaga attttcacag ctgtattcag gtaactgaag attttgtgtc tccagaacat
     1081 cttgtagagt catttcattt aacacaggaa ctgagacttt tggcagagaa cctctacttc
     1141 caatcgcacc atcatcacca ccatgattac aaggatgacg acgataagtg aggatccgaa
     1201 ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga
     1261 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1321 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1381 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1621 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1741 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     1861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     1981 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2041 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2101 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2161 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2221 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2281 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2341 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2401 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2461 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2521 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2581 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2641 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2701 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2761 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2821 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3181 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3241 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3301 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3361 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3421 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3481 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3541 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3601 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3661 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg
     3721 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3781 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3841 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3901 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3961 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     4021 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     4081 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     4141 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4201 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4261 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4321 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4381 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4441 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4501 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     4561 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4621 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4681 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     4741 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     4801 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4861 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4981 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     5041 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5101 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5161 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     5221 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5281 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5341 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5401 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5461 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5521 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5581 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5641 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5701 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5761 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5821 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5881 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5941 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     6001 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6061 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     6121 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     6181 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6241 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6301 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     6361 tctcgatccc gcgaaat
//
