LOCUS       62056_541        9314 bp DNA     circular SYN 14-MAY-2021
DEFINITION  Cloning vector CMV+ with onboard mCherry cassette, complete 
            sequence.
ACCESSION   MW987530
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9314)
  AUTHORS   Yang J, Lee J, Land MA, Lai S, Igoshin O, St-Pierre F.
  TITLE     A synthetic circuit for buffering gene dosage variation between 
            individual mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9314)
  AUTHORS   Yang J, Lee J, Land MA, Lai S, Igoshin O, St-Pierre F.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2021) Neuroscience, Baylor College of Medicine, 
            One Baylor Plaza, Suite S636, Houston, Texas 77030, United States
REFERENCE   3  (bases 1 to 9314)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2021) Neuroscience, Baylor College of Medicine, One Baylor 
            Plaza, Suite S636, Houston, Texas 77030, United States"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        163..360
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    361..379
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          588..1160
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1178..1894
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2338..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2846..2893
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2901..3920
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4053..4186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4223..4239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4247..4263)
                     /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(4271..4301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4316..4337)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4584..5762
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             5807..6514
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    6674..6729
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     rep_origin      complement(7106..7694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7868..8725)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8726..8830)
                     /label=AmpR promoter
     enhancer        9096..9314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
       61 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      121 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      181 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      241 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      301 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      361 tccctatcag tgatagagat cagatctccc tatcagtgat agagagctgt tttgacctcc
      421 atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt aagctgggta
      481 cccggggatc ctctagggcc tctgagctat tccagaagta gtgaagaggc ttttttggag
      541 gcctaggctt ttgcaaaaag ctccggatcg atcctgagaa cttcagggtg agtttgggga
      601 cccttgattg ttctttcttt ttcgctattg taaaattcat gttatatgga gggggcaaag
      661 ttttcagggt gttgtttaga atgggaagat gtcccttgta tcaccatgga ccctcatgat
      721 aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt attttctttt
      781 cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatt tttaaattca
      841 cttttgttta tttgtcagat tgtaagtact ttctctaatc actttttttt caaggcaatc
      901 agggtatatt atattgtact tcagcacagt tttagagaac aattgttata attaaatgat
      961 aaggtagaat atttctgcat ataaattctg gctggcgtgg aaatattctt attggtagaa
     1021 acaactacat cctggtcatc atcctgcctt tctctttatg gttacaatga tatacactgt
     1081 ttgagatgag gataaaatac tctgagtcca aaccgggccc ctctgctaac catgttcatg
     1141 ccttcttctt tttcctacag gtcctgcagg cgccaccatg gtgagcaagg gcgaggagct
     1201 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1261 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1321 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1381 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1441 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1501 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1561 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1621 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1681 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1741 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1801 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1861 cgggatcact ctcggcatgg acgagctgta caagtaggcg gccgcaatca acctctggat
     1921 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1981 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2041 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2101 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2161 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2221 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2281 tccgtggtgt tgtcctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     2341 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2401 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2461 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2521 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2581 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2641 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2701 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2761 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2821 attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa
     2881 gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     2941 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     3001 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     3061 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     3121 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     3181 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     3241 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     3301 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     3361 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     3421 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     3481 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     3541 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     3601 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     3661 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     3721 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     3781 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     3841 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     3901 ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct
     3961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4021 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4081 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4141 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4201 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4261 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4321 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4381 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4441 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4561 agaacatgta aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
     4621 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     4681 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     4741 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     4801 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     4861 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     4921 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     4981 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     5041 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     5101 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     5161 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     5221 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     5281 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     5341 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     5401 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     5461 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     5521 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     5581 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     5641 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     5701 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     5761 gaggtaccgg atcctctaga gtcgactccg gacgtcgacg gccaccatgg tgagcaaggg
     5821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     5881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     5941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     6001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     6061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     6121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     6181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     6241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     6301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     6361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     6421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     6481 ccactccacc ggcggcatgg acgagctgta caagtaatga gcggccgctt cactcctcag
     6541 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     6601 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     6661 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     6721 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     6781 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     6841 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     6901 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     6961 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     7021 ccagtcatag ctgtccctct tctcacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8881 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8941 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     9001 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     9061 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     9121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     9181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     9241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     9301 gtatttacgg taaa
//
