LOCUS       62056_6175        6145 bp DNA     circular SYN 18-OCT-2021
DEFINITION  Cloning vector pcDNA5_EGFP UBC, complete sequence.
ACCESSION   MW987535
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  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  JOURNAL   Unpublished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 6145)
  AUTHORS   Yang J, Lee J, Land MA, Lai S, Igoshin O, St-Pierre F.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2021) Neuroscience, Baylor College of Medicine, 
            One Baylor Plaza, Suite S636, Houston, Texas 77030, United States
REFERENCE   3  (bases 1 to 6145)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-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##
            On Oct 18, 2021 this sequence version replaced MW987535.1.
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    141..274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(311..327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(335..351)
                     /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(359..389)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(404..425)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(713..1301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1475..2332)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2333..2437)
                     /label=AmpR promoter
     promoter        2487..3698
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             3752..4468
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4571..4795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5079..5126
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             join(5134..6145,1..8)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg
       61 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
      121 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
      181 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      241 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
      301 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      361 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1141 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2461 ctgacgtcga cggatcggga gatctcggcc tccgcgccgg gttttggcgc ctcccgcggg
     2521 cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga
     2581 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     2641 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     2701 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     2761 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     2821 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     2881 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     2941 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     3001 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     3061 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     3121 gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat
     3181 gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     3241 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     3301 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     3361 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     3421 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     3481 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     3541 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     3601 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt
     3661 ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct gcaggtcgac
     3721 tctagaggat ccccgggtac cggtcgccac catggtgagc aagggcgagg agctgttcac
     3781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4441 cactctcggc atggacgagc tgtacaagta gggatccact agtccagtgt ggtggaattc
     4501 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     4561 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4621 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4681 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4741 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4801 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5041 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc
     5101 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct
     5161 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     5221 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     5281 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     5341 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     5401 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     5461 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     5521 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     5581 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     5641 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     5701 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     5761 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     5821 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     5881 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     5941 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     6001 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     6061 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     6121 cgacgcccca gcactcgtcc gaggg
//
