LOCUS       Exported                7457 bp DNA     circular SYN 31-AUG-2024
DEFINITION  Exported.
ACCESSION   V001259
VERSION     .
KEYWORDS    pCAG-T7pol
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  TITLE     Genetic Neuroengineering Group Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7457)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7457)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7457)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1742..4390
                     /codon_start=1
                     /label=T7 RNA polymerase
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYE
                     MGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQ
                     FLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFK
                     KNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIE
                     STGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGI
                     TGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANV
                     ITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLE
                     FMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYY
                     WLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFE
                     YAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAK
                     KVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVM
                     TLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVE
                     AMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLG
                     QFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHD
                     SFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLN
                     LRDILESDFAFA"
     primer_bind     complement(4479..4498)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4544..4599
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4598..4617)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(4960..4976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4984..5000)
                     /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)."
     promoter        complement(5008..5038)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5053..5074)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5132..5328
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    5334..5468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5536..5553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6469..7326)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7327..7431)
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccgccac
     1741 catgaacacg attaacatcg ctaagaacga cttctctgac atcgaactgg ctgctatccc
     1801 gttcaacact ctggctgacc attacggtga gcgtttagct cgcgaacagt tggcccttga
     1861 gcatgagtct tacgagatgg gtgaagcacg cttccgcaag atgtttgagc gtcaacttaa
     1921 agctggtgag gttgcggata acgctgccgc caagcctctc atcactaccc tactccctaa
     1981 gatgattgca cgcatcaacg actggtttga ggaagtgaaa gctaagcgcg gcaagcgccc
     2041 gacagccttc cagttcctgc aagaaatcaa gccggaagcc gtagcgtaca tcaccattaa
     2101 gaccactctg gcttgcctaa ccagtgctga caatacaacc gttcaggctg tagcaagcgc
     2161 aatcggtcgg gccattgagg acgaggctcg cttcggtcgt atccgtgacc ttgaagctaa
     2221 gcacttcaag aaaaacgttg aggaacaact caacaagcgc gtagggcacg tctacaagaa
     2281 agcatttatg caagttgtcg aggctgacat gctctctaag ggtctactcg gtggcgaggc
     2341 gtggtcttcg tggcataagg aagactctat tcatgtagga gtacgctgca tcgagatgct
     2401 cattgagtca accggaatgg ttagcttaca ccgccaaaat gctggcgtag taggtcaaga
     2461 ctctgagact atcgaactcg cacctgaata cgctgaggct atcgcaaccc gtgcaggtgc
     2521 gctggctggc atctctccga tgttccaacc ttgcgtagtt cctcctaagc cgtggactgg
     2581 cattactggt ggtggctatt gggctaacgg tcgtcgtcct ctggcgctgg tgcgtactca
     2641 cagtaagaaa gcactgatgc gctacgaaga cgtttacatg cctgaggtgt acaaagcgat
     2701 taacattgcg caaaacaccg catggaaaat caacaagaaa gtcctagcgg tcgccaacgt
     2761 aatcaccaag tggaagcatt gtccggtcga ggacatccct gcgattgaac gtgaagaact
     2821 cccgatgaaa ccggaagaca tcgacatgaa tcctgaggct ctcaccgcgt ggaaacgtgc
     2881 tgccgctgct gtgtaccgca aggacaaggc tcgcaagtct cgccgtatca gccttgagtt
     2941 catgcttgag caagccaata agtttgctaa ccataaggcc atctggttcc cttacaacat
     3001 ggactggcgc ggtcgtgttt acgctgtgtc aatgttcaac ccgcaaggta acgatatgac
     3061 caaaggactg cttacgctgg cgaaaggtaa accaatcggt aaggaaggtt actactggct
     3121 gaaaatccac ggtgcaaact gtgcgggtgt cgataaggtt ccgttccctg agcgcatcaa
     3181 gttcattgag gaaaaccacg agaacatcat ggcttgcgct aagtctccac tggagaacac
     3241 ttggtgggct gagcaagatt ctccgttctg cttccttgcg ttctgctttg agtacgctgg
     3301 ggtacagcac cacggcctga gctataactg ctcccttccg ctggcgtttg acgggtcttg
     3361 ctctggcatc cagcacttct ccgcgatgct ccgagatgag gtaggtggtc gcgcggttaa
     3421 cttgcttcct agtgaaaccg ttcaggacat ctacgggatt gttgctaaga aagtcaacga
     3481 gattctacaa gcagacgcaa tcaatgggac cgataacgaa gtagttaccg tgaccgatga
     3541 gaacactggt gaaatctctg agaaagtcaa gctgggcact aaggcactgg ctggtcaatg
     3601 gctggcttac ggtgttactc gcagtgtgac taagcgttca gtcatgacgc tggcttacgg
     3661 gtccaaagag ttcggcttcc gtcaacaagt gctggaagat accattcagc cagctattga
     3721 ttccggcaag ggtctgatgt tcactcagcc gaatcaggct gctggataca tggctaagct
     3781 gatttgggaa tctgtgagcg tgacggtggt agctgcggtt gaagcaatga actggcttaa
     3841 gtctgctgct aagctgctgg ctgctgaggt caaagataag aagactggag agattcttcg
     3901 caagcgttgc gctgtgcatt gggtaactcc tgatggtttc cctgtgtggc aggaatacaa
     3961 gaagcctatt cagacgcgct tgaacctgat gttcctcggt cagttccgct tacagcctac
     4021 cattaacacc aacaaagata gcgagattga tgcacacaaa caggagtctg gtatcgctcc
     4081 taactttgta cacagccaag acggtagcca ccttcgtaag actgtagtgt gggcacacga
     4141 gaagtacgga atcgaatctt ttgcactgat tcacgactcc ttcggtacca ttccggctga
     4201 cgctgcgaac ctgttcaaag cagtgcgcga aactatggtt gacacatatg agtcttgtga
     4261 tgtactggct gatttctacg accagttcgc tgaccagttg cacgagtctc aattggacaa
     4321 aatgccagca cttccggcta aaggtaactt gaacctccgt gacatcttag agtcggactt
     4381 cgcgttcgcg taagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4441 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4501 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4561 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4621 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4681 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4741 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4801 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4861 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     4921 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4981 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5041 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5101 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     5161 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5221 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5281 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     5341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5461 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7081 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7441 ccgaaaagtg ccacctg
//
