LOCUS       Exported                8801 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pXCX.CMV.tTA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8801)
  AUTHORS   Lee YB, Cosgrave AS, Glover CP, Bienemann A, Heywood D, Hobson RJ, 
            Uney JB
  TITLE     Increased utility in the CNS of a powerful neuron-specific 
            tetracycline-regulatable adenoviral system developed using a 
            post-transcriptional enhancer.
  JOURNAL   J Gene Med. 2005 May;7(5):576-83.
  PUBMED    15580589
REFERENCE   2  (bases 1 to 8801)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8801
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    484..708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     697..714
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(818..1825)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     1733..1752
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1848..1864)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1849..1865)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1954..1973)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1955..1973)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2018..2221)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2047..2067)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2222..2601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             6353..6544
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6538..6560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6646..6786
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6701..6720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6801..6818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6972..7560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7052..7071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7731..8591)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8354..8373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8592..8696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8764..8782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cccatcatca taatatacct tattttggat tgaagccaat atgataatga gggggtggag
       61 tttgtgacgt ggcgcggggc gtgggaacgg ggcgggtgac gtagtagtgt ggcggaagtg
      121 tgatgttgca agtgtggcgg aacacatgta agcgccggat gtggtaaaag tgacgttttt
      181 ggtgtgcgcc ggtgtatacg ggaagtgaca attttcgcgc ggttttaggc ggatgttgta
      241 gtaaatttgg gcgtaaccaa gtaatatttg gccattttcg cgggaaaact gaataagagg
      301 aagtgaaatc tgaataattc tgtgttactc atagcgcgta atatttgtct agggccgcgg
      361 ggactttgac cgtttacgtg gagactcgcc caggtgtttt tctcaggtgt tttccgcgtt
      421 ccgggtcaaa gttggcgttt tattattata gtcagctcta gctggttctt tccgcctcag
      481 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
      541 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
      601 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
      661 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
      721 agcgagctct agatgcatgc tcgagcggcc gccagtgtga tggaatctgc agaattccag
      781 cacactggcg gccgttacta gtggatcctc gcgcccccta cccaccgtac tcgtcaattc
      841 caagggcatc ggtaaacatc tgctcaaact cgaagtcggc catatccaga gcgccgtagg
      901 gggcggagtc gtggggggta aatcccggac ccggggaatc cccgtccccc aacatgtcca
      961 gatcgaaatc gtctagcgcg tcggcatgcg ccatcgccac gtcctcgccg tctaagtgga
     1021 gctcgtcccc caggctgaca tcggtcgggg gggccgtcga cagtctgcgc gtgtgtcccg
     1081 cggggagaaa ggacaggcgc ggagccgcca gccccgcctc ttcgggggcg tcgtcgtccg
     1141 ggagatcgag caggccctcg atggtagacc cgtaattgtt tttcgtacgc gcgcggctgt
     1201 acgcggaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc
     1261 cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata
     1321 atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt
     1381 gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat
     1441 tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact
     1501 gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag
     1561 cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag
     1621 ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc
     1681 gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt
     1741 tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg ctgttaatca
     1801 ctttactttt atctaatcta gacatatgaa ttcgatatca agcttatcga taccgtcgac
     1861 ctcgagcggc cgccagtgtg atggaatctg cagaattcca gcacactggc ggccgttact
     1921 agtggatccg agctcggtac caagcttggg tctccctata gtgagtcgta ttaatttcga
     1981 taagccagta agcagtgggt tctctagtta gccagagagc tctgcttata tagacctccc
     2041 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     2101 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     2161 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     2221 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     2281 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     2341 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     2401 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     2461 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     2521 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     2581 aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcta gagtggaagg
     2641 tgctgaggta cgatgagacc cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata
     2701 ttaggaacca gcctgtgatg ctggatgtga ccgaggagct gaggcccgat cacttggtgc
     2761 tggcctgcac ccgcgctgag tttggctcta gcgatgaaga tacagattga ggtactgaaa
     2821 tgtgtgggcg tggcttaagg gtgggaaaga atatataagg tgggggtctt atgtagtttt
     2881 gtatctgttt tgcagcagcc gccgccgcca tgagcaccaa ctcgtttgat ggaagcattg
     2941 tgagctcata tttgacaacg cgcatgcccc catgggccgg ggtgcgtcag aatgtgatgg
     3001 gctccagcat tgatggtcgc cccgtcctgc ccgcaaactc tactaccttg acctacgaga
     3061 ccgtgtctgg aacgccgttg gagactgcag cctccgccgc cgcttcagcc gctgcagcca
     3121 ccgcccgcgg gattgtgact gactttgctt tcctgagccc gcttgcaagc agtgcagctt
     3181 cccgttcatc cgcccgcgat gacaagttga cggctctttt ggcacaattg gattctttga
     3241 cccgggaact taatgtcgtt tctcagcagc tgttggatct gcgccagcag gtttctgccc
     3301 tgaaggcttc ctcccctccc aatgcggttt aaaacataaa taaaaaacca gactctgttt
     3361 ggatttggat caagcaagtg tcttgctgtc tttatttagg ggttttgcgc gcgcggtagg
     3421 cccgggacca gcggtctcgg tcgttgaggg tcctgtgtat tttttccagg acgtggtaaa
     3481 ggtgactctg gatgttcaga tacatgggca taagcccgtc tctggggtgg aggtagcacc
     3541 actgcagagc ttcatgctgc ggggtggtgt tgtagatgat ccagtcgtag caggagcgct
     3601 gggcgtggtg cctaaaaatg tctttcagta gcaagctgat tgccaggggc aggcccttgg
     3661 tgtaagtgtt tacaaagcgg ttaagctggg atgggtgcat acgtggggat atgagatgca
     3721 tcttggactg tatttttagg ttggctatgt tcccagccat atccctccgg ggattcatgt
     3781 tgtgcagaac caccagcaca gtgtatccgg tgcacttggg aaatttgtca tgtagcttag
     3841 aaggaaatgc gtggaagaac ttggagacgc ccttgtgacc tccaagattt tccatgcatt
     3901 cgtccataat gatggcaatg ggcccacggg cggcggcctg ggcgaagata tttctgggat
     3961 cactaacgtc atagttgtgt tccaggatga gatcgtcata ggccattttt acaaagcgcg
     4021 ggcggagggt gccagactgc ggtataatgg ttccatccgg cccaggggcg tagttaccct
     4081 cacagatttg catttcccac gctttgagtt cagatggggg gatcatgtct acctgcgggg
     4141 cgatgaagaa aacggtttcc ggggtagggg agatcagctg ggaagaaagc aggttcctga
     4201 gcagctgcga cttaccgcag ccggtgggcc cgtaaatcac acctattacc gggtgcaact
     4261 ggtagttaag agagctgcag ctgccgtcat ccctgagcag gggggccact tcgttaagca
     4321 tgtccctgac tcgcatgttt tccctgacca aatccgccag aaggcgctcg ccgcccagcg
     4381 atagcagttc ttgcaaggaa gcaaagtttt tcaacggttt gagaccgtcc gccgtaggca
     4441 tgcttttgag cgtttgacca agcagttcca ggcggtccca cagctcggtc acctgctcta
     4501 cggcatctcg atccagcata tctcctcgtt tcgcgggttg gggcggcttt cgctgtacgg
     4561 cagtagtcgg tgctcgtcca gacgggccag ggtcatgtct ttccacgggc gcagggtcct
     4621 cgtcagcgta gtctgggtca cggtgaaggg gtgcgctccg ggctgcgcgc tggccagggt
     4681 gcgcttgagg ctggtcctgc tggtgctgaa gcgctgccgg tcttcgccct gcgcgtcggc
     4741 caggtagcat ttgaccatgg tgtcatagtc cagcccctcc gcggcgtggc ccttggcgcg
     4801 cagcttgccc ttggaggagg cgccgcacga ggggcagtgc agacttttga gggcgtagag
     4861 cttgggcgcg agaaataccg attccgggga gtaggcatcc gcgccgcagg ccccgcagac
     4921 ggtctcgcat tccacgagcc aggtgagctc tggccgttcg gggtcaaaaa ccaggtttcc
     4981 cccatgcttt ttgatgcgtt tcttacctct ggtttccatg agccggtgtc cacgctcggt
     5041 gacgaaaagg ctgtccgtgt ccccgtatac agacttgaga ggcctgtcct cgaccgatgc
     5101 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     5161 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     5221 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     5281 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     5341 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     5401 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     5461 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     5521 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc
     5581 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     5641 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     5701 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     5761 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     5821 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     5881 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     5941 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     6001 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     6061 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     6121 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     6181 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     6241 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     6301 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     6361 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     6421 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     6481 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     6541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     6661 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     6721 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6781 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6841 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6901 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7381 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8041 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8341 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8701 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8761 aaaaataggc gtatcacgag gccctttcgt cttcaagaat t
//
