LOCUS       Exported                6189 bp ds-DNA     circular SYN 18-OCT-2013
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6189)
  AUTHORS   Schwarz
  TITLE     Direct Submission
  JOURNAL   Exported Sep 18, 2018 from SnapGene 4.2.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6189
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="This feature has 2 segments:
                        1: 235 .. 484 / #ffffff
                        2: 485 .. 614 / #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     769..789
                     /label=CMV_fwd
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     CDS             863..1579
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  863 ..  865 / #e1ff2f
                        2:  866 ..  868 / #e1ff2f / 1a
                        3:  869 .. 1579 / #e1ff2f"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(908..929)
                     /label=EGFP-N
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     1516..1537
                     /label=EGFP-C
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     misc_feature    1580..1645
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     primer_bind     1652..1677
                     /label=DHFR_5'
                     /note="color: black; sequence: 
                     ACTCTAGAATCAGTCTGATTGCGGCGTT"
     CDS             1658..2131
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /note="color: #993366"
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     primer_bind     complement(2116..2131)
                     /label=agt_DHFR_3'
                     /note="color: black; sequence: AGTCCGCCGCTCCAGAATC"
     CDS             2132..2161
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     primer_bind     complement(2196..2213)
                     /label=BGH_rev
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    2202..2426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      2472..2900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2914..3244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3095..3230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        3292..3339
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3358..3732
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="color: #ccffcc"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3862..3983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4032..4048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(4032..4048)
                     /label=M13_rev
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     protein_bind    4056..4072
                     /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)."
                     /note="color: #31849b"
     promoter        complement(4080..4110)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4080 .. 4086 / #ffffff / -10
                        2: 4087 .. 4104 / #ffffff
                        3: 4105 .. 4110 / #ffffff / -35"
     protein_bind    4125..4146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4434..5022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5193..6053)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5193 .. 5984 / #ccffcc
                        2: 5985 .. 6053 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6054..6158)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1021 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc
     1081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1441 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     1501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1561 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt
     1621 cgacggtacc gcgggcccgg gatccaccgg atctagaatc agtctgattg cggcgttagc
     1681 ggtagatcgc gttatcggca tggaaaacgc catgccgtgg aacctgcctg ccgatctcgc
     1741 ctggtttaaa cgcaacacct taaataaacc cgtgattatg ggccgccata cctgggaatc
     1801 aatcggtcgt ccgttgccag gacgcaaaaa tattatcctc agcagtcaac cgggtacgga
     1861 cgatcgcgta acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg gtgacgtacc
     1921 agaaatcatg gtgattggcg gcggtcgcgt ttatgaacag ttcttgccaa aagcgcaaaa
     1981 actgtatctg acgcatatcg acgcagaagt ggaaggcgac acccatttcc cggattacga
     2041 gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactctca
     2101 cagctattgc tttgagattc tggagcggcg ggaacaaaaa ctcatctcag aagaggatct
     2161 gtagactaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2221 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2281 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2341 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2401 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2461 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2521 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2581 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     2641 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2701 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2761 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2821 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     2881 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2941 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3061 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3121 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3181 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3241 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     3301 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     3361 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     3421 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     3481 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     3541 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     3601 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     3661 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     3721 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3781 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3841 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3961 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4021 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4081 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4141 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4201 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4621 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4861 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6181 cctgacgtc
//
