LOCUS       40924_18531        8584 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETDuet-hTERT-noMCS2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8584)
  AUTHORS   Hansen DT, Thiyagarajan T, Larson AC, Hansen JL.
  TITLE     Telomerase repeat amplification protocol (TRAP) activity upon 
            recombinant expression and purification of human telomerase in a 
            bacterial system
  JOURNAL   Protein Expr. Purif. (2016) In press
  PUBMED    26965413
REFERENCE   2  (bases 1 to 8584)
  AUTHORS   Hansen DT, Hansen JL.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2015) Department of Biochemistry and Molecular 
            Biology, Medical University of South Carolina, 173 Ashley Avenue, 
            Charleston, SC 29425, USA
REFERENCE   3  (bases 1 to 8584)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8584)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr. Purif. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-DEC-2015) Department of Biochemistry and Molecular Biology, 
            Medical University of South Carolina, 173 Ashley Avenue, Charleston,
            SC 29425, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /label=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             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             72..3467
                     /codon_start=1
                     /label=hTERT
                     /note="human telomerase reverse transcriptase, the
                     catalytic subunit of telomerase"
                     /translation="MPRAPRCRAVRSLLRSHYREVLPLATFVRRLGPQGWRLVQRGDPA
                     AFRALVAQCLVCVPWDARPPPAAPSFRQVSCLKELVARVLQRLCERGAKNVLAFGFALL
                     DGARGGPPEAFTTSVRSYLPNTVTDALRGSGAWGLLLRRVGDDVLVHLLARCALFVLVA
                     PSCAYQVCGPPLYQLGAATQARPPPHASGPRRRLGCERAWNHSVREAGVPLGLPAPGAR
                     RRGGSASRSLPLPKRPRRGAAPEPERTPVGQGSWAHPGRTRGPSDRGFCVVSPARPAEE
                     ATSLEGALSGTRHSHPSVGRQHHAGPPSTSRPPRPWDTPCPPVYAETKHFLYSSGDKEQ
                     LRPSFLLSSLRPSLTGARRLVETIFLGSRPWMPGTPRRLPRLPQRYWQMRPLFLELLGN
                     HAQCPYGVLLKTHCPLRAAVTPAAGVCAREKPQGSVAAPEEEDTDPRRLVQLLRQHSSP
                     WQVYGFVRACLRRLVPPGLWGSRHNERRFLRNTKKFISLGKHAKLSLQELTWKMSVRDC
                     AWLRRSPGVGCVPAAEHRLREEILAKFLHWLMSVYVVELLRSFFYVTETTFQKNRLFFY
                     RKSVWSKLQSIGIRQHLKRVQLRELSEAEVRQHREARPALLTSRLRFIPKPDGLRPIVN
                     MDYVVGARTFRREKRAERLTSRVKALFSVLNYERARRPGLLGASVLGLDDIHRAWRTFV
                     LRVRAQDPPPELYFVKVDVTGAYDTIPQDRLTEVIASIIKPQNTYCVRRYAVVQKAAHG
                     HVRKAFKSHVSTLTDLQPYMRQFVAHLQETSPLRDAVVIEQSSSLNEASSGLFDVFLRF
                     MCHHAVRIRGKSYVQCQGIPQGSILSTLLCSLCYGDMENKLFAGIRRDGLLLRLVDDFL
                     LVTPHLTHAKTFLRTLVRGVPEYGCVVNLRKTVVNFPVEDEALGGTAFVQMPAHGLFPW
                     CGLLLDTRTLEVQSDYSSYARTSIRASLTFNRGFKAGRNMRRKLFGVLRLKCHSLFLDL
                     QVNSLQTVCTNIYKILLLQAYRFHACVLQLPFHQQVWKNPTFFLRVISDTASLCYSILK
                     AKNAGMSLGAKGAAGPLPSEAVQWLCHQAFLLKLTRHRVTYVPLLGSLRTAQTQLSRKL
                     PGTTLTALEAAANPALPSDFKTILD"
     CDS             3530..3574
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      3626..3673
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3710..4165
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4283..5140)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5141..5233)
                     /label=AmpR promoter
     rep_origin      5314..5902
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(6088..6230)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6335..6523)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(7061..7082)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(7098..8177)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(8178..8255)
                     /label=lacI promoter
     promoter        8568..8584
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc tatgccgcgc gctccccgct gccgagccgt gcgctccctg ctgcgcagcc
      121 actaccgcga ggtgctgccg ctggccacgt tcgtgcggcg cctggggccc cagggctggc
      181 ggctggtgca gcgcggggac ccggcggctt tccgcgcgct ggtggcccag tgcctggtgt
      241 gcgtgccctg ggacgcacgg ccgccccccg ccgccccctc cttccgccag gtgtcctgcc
      301 tgaaggagct ggtggcccga gtgctgcaga ggctgtgcga gcgcggcgcg aagaacgtgc
      361 tggccttcgg cttcgcgctg ctggacgggg cccgcggggg cccccccgag gccttcacca
      421 ccagcgtgcg cagctacctg cccaacacgg tgaccgacgc actgcggggg agcggggcgt
      481 gggggctgct gctgcgccgc gtgggcgacg acgtgctggt tcacctgctg gcacgctgcg
      541 cgctctttgt gctggtggct cccagctgcg cctaccaggt gtgcgggccg ccgctgtacc
      601 agctcggcgc tgccactcag gcccggcccc cgccacacgc tagtggaccc cgaaggcgtc
      661 tgggatgcga acgggcctgg aaccatagcg tcagggaggc cggggtcccc ctgggcctgc
      721 cagccccggg tgcgaggagg cgcgggggca gtgccagccg aagtctgccg ttgcccaaga
      781 ggcccaggcg tggcgctgcc cctgagccgg agcggacgcc cgttgggcag gggtcctggg
      841 cccacccggg caggacgcgt ggaccgagtg accgtggttt ctgtgtggtg tcacctgcca
      901 gacccgccga agaagccacc tctttggagg gtgcgctctc tggcacgcgc cactcccacc
      961 catccgtggg ccgccagcac cacgcgggcc ccccatccac atcgcggcca ccacgtccct
     1021 gggacacgcc ttgtcccccg gtgtacgccg agaccaagca cttcctctac tcctcaggcg
     1081 acaaggagca gctgcggccc tccttcctac tcagctctct gaggcccagc ctgactggcg
     1141 ctcggaggct cgtggagacc atctttctgg gttccaggcc ctggatgcca gggactcccc
     1201 gcaggttgcc ccgcctgccc cagcgctact ggcaaatgcg gcccctgttt ctggagctgc
     1261 ttgggaacca cgcgcagtgc ccctacgggg tgctcctcaa gacgcactgc ccgctgcgag
     1321 ctgcggtcac cccagcagcc ggtgtctgtg cccgggagaa gccccagggc tctgtggcgg
     1381 cccccgagga ggaggacaca gacccccgtc gcctggtgca gctgctccgc cagcacagca
     1441 gcccctggca ggtgtacggc ttcgtgcggg cctgcctgcg ccggctggtg cccccaggcc
     1501 tctggggctc caggcacaac gaacgccgct tcctcaggaa caccaagaag ttcatctccc
     1561 tggggaagca tgccaagctc tcgctgcagg agctgacgtg gaagatgagc gtgcgggact
     1621 gcgcttggct gcgcaggagc ccaggggttg gctgtgttcc ggccgcagag caccgtctgc
     1681 gtgaggagat cctggccaag ttcctgcact ggctgatgag tgtgtacgtc gtcgagctgc
     1741 tcaggtcttt cttttatgtc acggagacca cgtttcaaaa gaacaggctc tttttctacc
     1801 ggaagagtgt ctggagcaag ttgcaaagca ttggaatcag acagcacttg aagagggtgc
     1861 agctgcggga gctgtcggaa gcagaggtca ggcagcatcg ggaagccagg cccgccctgc
     1921 tgacgtccag actccgcttc atccccaagc ctgacgggct gcggccgatt gtgaacatgg
     1981 actacgtcgt gggagccaga acgttccgca gagaaaagag ggccgagcgt ctcacctcga
     2041 gggtgaaggc actgttcagc gtgctcaact acgagcgggc gcggcgcccc ggcctcctgg
     2101 gcgcctctgt gctgggcctg gacgatatcc acagggcctg gcgcaccttc gtgctgcgtg
     2161 tgcgggccca ggacccgccg cctgagctgt actttgtcaa ggtggatgtg acgggcgcgt
     2221 acgacaccat cccccaggac aggctcacgg aggtcatcgc cagcatcatc aaaccccaga
     2281 acacgtactg cgtgcgtcgg tatgccgtgg tccagaaggc cgcccatggg cacgtccgca
     2341 aggccttcaa gagccacgtc tctaccttga cagacctcca gccgtacatg cgacagttcg
     2401 tggctcacct gcaggagacc agcccgctga gggatgccgt cgtcatcgag cagagctcct
     2461 ccctgaatga ggccagcagt ggcctcttcg acgtcttcct acgcttcatg tgccaccacg
     2521 ccgtgcgcat caggggcaag tcctacgtcc agtgccaggg gatcccgcag ggctccatcc
     2581 tctccacgct gctctgcagc ctgtgctacg gcgacatgga gaacaagctg tttgcgggga
     2641 ttcggcggga cgggctgctc ctgcgtttgg tggatgattt cttgttggtg acacctcacc
     2701 tcacccacgc gaaaaccttc ctcaggaccc tggtccgagg tgtccctgag tatggctgcg
     2761 tggtgaactt gcggaagaca gtggtgaact tccctgtaga agacgaggcc ctgggtggca
     2821 cggcttttgt tcagatgccg gcccacggcc tattcccctg gtgcggcctg ctgctggata
     2881 cccggaccct ggaggtgcag agcgactact ccagctatgc ccggacctcc atcagagcca
     2941 gtctcacctt caaccgcggc ttcaaggctg ggaggaacat gcgtcgcaaa ctctttgggg
     3001 tcttgcggct gaagtgtcac agcctgtttc tggatttgca ggtgaacagc ctccagacgg
     3061 tgtgcaccaa catctacaag atcctcctgc tgcaggcgta caggtttcac gcatgtgtgc
     3121 tgcagctccc atttcatcag caagtttgga agaaccccac atttttcctg cgcgtcatct
     3181 ctgacacggc ctccctctgc tactccatcc tgaaagccaa gaacgcaggg atgtcgctgg
     3241 gggccaaggg cgccgccggc cctctgccct ccgaggccgt gcagtggctg tgccaccaag
     3301 cattcctgct caagctgact cgacaccgtg tcacctacgt gccactcctg gggtcactca
     3361 ggacagccca gacgcagctg agtcggaagc tcccggggac gacgctgact gccctggagg
     3421 ccgcagccaa cccggcactg ccctcagact tcaagaccat cctggactga gcggccgcat
     3481 aatgcttaag tcgaacagaa agtaatcgta ttgtaccctc gagtctggta aagaaaccgc
     3541 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     3601 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3661 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     3721 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3781 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3841 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3901 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3961 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4021 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4081 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4141 aacaaaatat taacgtttac aatttctggc ggcacgatgg catgagatta tcaaaaagga
     4201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat
     5161 gattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5221 aaaaataaac aaataggtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5461 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6061 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     6121 cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6181 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     6241 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6301 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6361 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     6421 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     6481 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     6541 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     6601 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     6661 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     6721 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     6781 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     6841 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     6901 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     6961 acgacaggag cacgatcatg ctagtcatgc cccgcgccca ccggaaggag ctgactgggt
     7021 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     7081 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7141 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     7201 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     7261 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     7321 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     7381 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     7441 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     7501 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     7561 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     7621 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     7681 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     7741 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     7801 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     7861 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     7921 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     7981 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     8041 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     8101 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     8161 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     8221 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     8281 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     8341 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     8401 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     8461 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     8521 gatgcgtccg gcgtagagga tcgagatcga tctcgatccc gcgaaattaa tacgactcac
     8581 tata
//
