LOCUS       Exported                6787 bp ds-DNA    circular SYN 04-NOV-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pET28a Cdk2ap1-Delta-N (MT2B2)-MutTER
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6787)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Nov 4, 2021 from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6787
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(50..865)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      987..1575
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1761..1903
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2005..2196)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3005..4087)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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"
     promoter        complement(4088..4165)
                     /gene="lacI"
                     /note="lacI promoter"
                     /note="
                     "
     promoter        4474..4492
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4493..4517
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     RBS             4532..4554
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4573..4590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             4600..4617
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     gene            4621..5721
                     /gene="MBP"
                     /locus_tag="MBP"
                     /label="MBP"
                     /note="MBP"
     gene            5737..5757
                     /gene="TEV"
                     /locus_tag="TEV"
                     /label="TEV"
                     /note="TEV"
     CDS             5737..5757
                     /gene="CDS"
                     /locus_tag="TEV site(1)"
                     /label="TEV site(1)"
                     /note="TEV"
                     /note="CDS"
     CDS             5809..5829
                     /gene="CDS"
                     /locus_tag="TEV site"
                     /label="TEV site"
                     /note="TRS"
                     /note="CDS"
     CDS             5839..6075
                     /codon_start=1
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /product="cyclin-dependent kinase 2-associated protein 1"
                     /note="Cdk2ap1"
                     /note="CDK2-associated protein 1; putative oral cancer 
                     suppressor; deleted in oral cancer 1"
                     /db_xref="GI:49355794"
                     /db_xref="CCDS:CCDS39279.1"
                     /db_xref="GeneID:13445"
                     /db_xref="MGI:MGI:1202069"
                     /protein_id="NP_038840.2"
                     /translation="MATSSQYRQLLSDYGPPSLGYTQGTGNSQVPQSKYAELLAIIEEL
                     GKEIRPTYAGSKSAMERLKRGIIHARSLVRECLA"
     exon            5839..5907
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /inference="alignment:same species:1.39.8"
     attenuator      5839..5850
                     /note="NLS"
     exon            5908..6034
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /inference="alignment:same species:1.39.8"
     exon            6035..6075
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /inference="alignment:same species:1.39.8"
     exon            6076
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /inference="alignment:same species:1.39.8"
     CDS             6076
                     /codon_start=1
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /product="cyclin-dependent kinase 2-associated protein 1"
                     /note="Cdk2ap1"
                     /note="CDK2-associated protein 1; putative oral cancer 
                     suppressor; deleted in oral cancer 1"
                     /db_xref="GI:49355794"
                     /db_xref="CCDS:CCDS39279.1"
                     /db_xref="GeneID:13445"
                     /db_xref="MGI:MGI:1202069"
                     /protein_id="NP_038840.2"
                     /translation=""
     exon            6090..6099
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /inference="alignment:same species:1.39.8"
     CDS             6090..6099
                     /codon_start=1
                     /gene="Cdk2ap1"
                     /gene_synonym="Apc10; Cdkap1; doc-1; Doc1; DORC1; p12; 
                     ST19"
                     /product="cyclin-dependent kinase 2-associated protein 1"
                     /note="Cdk2ap1"
                     /note="CDK2-associated protein 1; putative oral cancer 
                     suppressor; deleted in oral cancer 1"
                     /db_xref="GI:49355794"
                     /db_xref="CCDS:CCDS39279.1"
                     /db_xref="GeneID:13445"
                     /db_xref="MGI:MGI:1202069"
                     /protein_id="NP_038840.2"
                     /translation="ARS"
     CDS             6121..6138
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      6205..6252
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6289..6744
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
       61 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
      121 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
      181 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
      241 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
      301 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
      361 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
      421 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
      481 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
      541 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
      601 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
      661 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
      721 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
      781 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
      841 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
      901 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     1141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1621 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1681 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1741 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1801 tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     1861 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     1921 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1981 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2041 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     2101 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     2161 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     2221 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     2281 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     2341 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     2401 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     2461 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     2521 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     2581 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     2641 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     2701 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     2761 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     2821 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     2881 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     2941 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     3001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     3121 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     3181 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     3241 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     3301 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     3361 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     3421 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     3481 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     3541 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     3601 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     3661 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     3721 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     3781 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     3841 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     3901 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     3961 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     4021 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     4081 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     4141 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     4201 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     4261 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     4321 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     4381 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     4441 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     4501 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
     4561 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
     4621 atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
     4681 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
     4741 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
     4801 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
     4861 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
     4921 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
     4981 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
     5041 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
     5101 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
     5161 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
     5221 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
     5281 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
     5341 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
     5401 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
     5461 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
     5521 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
     5581 accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     5641 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     5701 gccctgaaag acgcgcagac taattcgaat ttcgaggaaa atttatattt tcaaagcggc
     5761 accgtggatg ccggcgccga tgccggcaag ccccggcccg gcagcagcga gaacctgtac
     5821 ttccagggcg gccggccgat ggcgacctcc tcccaatatc gccagctgct gagtgactat
     5881 gggcctccat cgctaggcta cacccaggga actggaaata gccaggtgcc tcagagtaaa
     5941 tatgcagagc tgctggccat cattgaagag ttggggaaag agatcagacc gacgtatgca
     6001 gggagcaaga gcgccatgga gaggctaaaa cgaggcatca ttcatgcccg gagcctggtt
     6061 cgggagtgct tggctgaagc ggcagcgaat gccaggtcct gatgagcggc cgcactcgag
     6121 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     6181 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     6241 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     6301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6361 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6421 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6481 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6601 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6661 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6721 acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6781 ctatttg
//
