pcDNA3-CFP

LOCUS       pcDNA3-CFP             6159 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Mammalian expression vector for cloning your gene fused to CFP.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3-CFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  TITLE     Fluorescent Protein Plasmids
REFERENCE   2  (bases 1 to 6159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6159
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             965..1684
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1010..1031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1271..1290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1618..1639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1700..1718)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1701..1718)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1738..1755)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1744..1968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2014..2442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2101..2120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2311..2332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2451..2471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2456..2785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2636..2771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2698..2717
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2852..3646
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2906..2925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3516..3535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3820..3941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3857..3876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3911..3930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3990..4006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3990..4006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4003..4025)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4014..4030
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(4038..4068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4083..4104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4221..4238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4392..4980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4472..4491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5151..6011)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5774..5793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6012..6116)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
      901 actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct
      961 cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1021 ggacggcgac gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac
     1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1141 caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat
     1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat
     1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1381 gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa
     1441 gaacggcatc aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1561 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1681 gtaatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     1741 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1801 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1861 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1921 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1981 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2041 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2101 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2161 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2221 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2281 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2341 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2401 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2701 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2761 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     2821 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2881 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2941 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3001 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3061 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3121 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3181 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3241 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3301 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3361 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3421 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3481 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3541 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3601 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3661 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3721 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3781 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3961 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4021 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4081 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4141 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4201 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5281 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6121 tttccccgaa aagtgccacc tgacgtcgac ggatcggga
//