LOCUS       Stim1(233-685)(s        6209 bp DNA     circular SYN 27-AUG-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    mCerulean N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6209)
  AUTHORS   Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS
  TITLE     Cerulean, Venus, and VenusY67C FRET reference standards.
  JOURNAL   Biophys J. 2006 Dec 15. 91(12):L99-L101.
  PUBMED    17040988
REFERENCE   2  (bases 1 to 6209)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 2024 from SnapGene 6.1.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biophys J. 
            2006 Dec 15. 91(12):L99-L101."
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     CDS             622..657
                     /codon_start=1
                     /label=S1(233-685)(ssra)
                     /note="This feature has 2 segments:
                        1: 622 .. 645 / #00ff00 / ssra-wt
                        2: 646 .. 657 / #ffff00 / linker"
                     /translation="AANDENYAGGPG"
     CDS             658..2067
                     /codon_start=1
                     /label=S1(233-685)(ssra)
                     /note="This feature has 5 segments:
                        1:  658 ..  990 / #ff99cc
                        2:  991 .. 1005 / #ffff00 / linker
                        3: 1006 .. 1029 / #00ff00 / ssra-wt
                        4: 1030 .. 1041 / #ffff00 / linker
                        5: 1042 .. 2067 / #ff99cc"
                     /translation="QNRYSKEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEV
                     EKVHLEKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELE
                     SHSSWYAGSGGGAANDENYAGGPGPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKE
                     GAEKIKKKRNTLFGTFHVAHSSSLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILC
                     GFQIVNNPGIHSLVAALNIDPSWMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQ
                     SSVRQRLTEPQHGLGSQRDLTHSDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNG
                     SHRLIEGVHPGSLVEKLPDSPALAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSS
                     RSHSPSSPDPDTPSPVGDSRALQASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKK
                     FPLKIFKKPLKK"
     CDS             2068..2103
                     /codon_start=1
                     /label=S1(233-685)(ssra)
                     /note="This feature has 2 segments:
                        1: 2068 .. 2079 / #ffff00 / linker
                        2: 2080 .. 2103 / #00ff00 / ssra-wt"
                     /translation="SGGGACNDENYA"
     regulatory      2149..2158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             2155..2874
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2155 .. 2157 / #00ffff
                        2: 2158 .. 2160 / #00ffff / 1a
                        3: 2161 .. 2874 / #00ffff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(2461..2480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     2808..2829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     polyA_signal    2997..3118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3034..3053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     3088..3107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      complement(3125..3580)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(3262..3283)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     primer_bind     3474..3493
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     promoter        3607..3711
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3713..4070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(3736..3756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /note="color: black; sequence: ACCCTAACTGACACACATTCC"
     rep_origin      3921..4056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     3983..4002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(4006..4017)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     CDS             4105..4899
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4159..4178)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
                     /note="color: black; sequence: GCCCAGTCATAGCCGAATAG"
     primer_bind     4769..4788
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
                     /note="color: black; sequence: CGTTGGCTACCCGTGATATT"
     primer_bind     complement(5087..5106)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
                     /note="color: black; sequence: TTGTCTCCTTCCGTGTTTCA"
     polyA_signal    5131..5178
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      5507..6095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     5996..6015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat ggcagccaat gatgagaact acgctggagg gcccggacag
      661 aaccgttact ccaaggagca catgaagaag atgatgaagg acttggaggg gttacaccga
      721 gctgagcaga gtctgcatga ccttcaggaa aggctgcaca aggcccagga ggagcaccgc
      781 acagtggagg tggagaaggt ccatctggaa aagaagctgc gcgatgagat caaccttgct
      841 aagcaggaag cccagcggct gaaggagctg cgggagggta ctgagaatga gcggagccgc
      901 caaaaatatg ctgaggagga gttggagcag gttcgggagg ccttgaggaa agcagagaag
      961 gagctagaat ctcacagctc atggtatgct ggatccggag gtggagcagc caatgatgag
     1021 aactacgctg gagggcccgg accagaggcc cttcagaagt ggctgcagct gacacatgag
     1081 gtggaggtgc aatattacaa catcaagaag caaaatgctg agaagcagct gctggtggcc
     1141 aaggaggggg ctgagaagat aaaaaagaag agaaacacac tctttggcac cttccacgtg
     1201 gcccacagct cttccctgga tgatgtagat cataaaattc taacagctaa gcaagcactg
     1261 agcgaggtga cagcagcatt gcgggagcgc ctgcaccgct ggcaacagat cgagatcctc
     1321 tgtggcttcc agattgtcaa caaccctggc atccactcac tggtggctgc cctcaacata
     1381 gaccccagct ggatgggcag tacacgcccc aaccctgctc acttcatcat gactgacgac
     1441 gtggatgaca tggatgagga gattgtgtct cccttgtcca tgcagtcccc tagcctgcag
     1501 agcagtgttc ggcagcgcct gacggagcca cagcatggcc tgggatctca gagggatttg
     1561 acccattccg attcggagtc ctccctccac atgagtgacc gccagcgtgt ggcccccaaa
     1621 cctcctcaga tgagccgtgc tgcagacgag gctctcaatg ccatgacttc caatggcagc
     1681 caccggctga tcgagggggt ccacccaggg tctctggtgg agaaactgcc tgacagccct
     1741 gccctggcca agaaggcatt actggcgctg aaccatgggc tggacaaggc ccacagcctg
     1801 atggagctga gcccctcagc cccacctggt ggctctccac atttggattc ttcccgttct
     1861 cacagcccca gctccccaga cccagacaca ccatctccag ttggggacag ccgagccctg
     1921 caagccagcc gaaacacacg cattccccac ctggctggca agaaggctgt ggctgaggag
     1981 gataatggct ctattggcga ggaaacagac tccagcccag gccggaagaa gtttcccctc
     2041 aaaatcttta agaagcctct taagaagtcc ggaggtggag catgcaatga tgagaactac
     2101 gctcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg
     2161 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2281 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccctga cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2461 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaacg ccatcagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc
     2641 aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2821 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     2881 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2941 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3001 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3061 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3121 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3181 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3241 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3301 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3361 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3421 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3481 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3541 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3601 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3661 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3721 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3841 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3901 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3961 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4021 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4081 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4141 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4201 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4261 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4321 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4381 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4441 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4501 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4561 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4621 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4681 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4741 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4801 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4861 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4921 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5041 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5101 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5161 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5221 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5281 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5341 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5401 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5461 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5521 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5581 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5641 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5701 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5761 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5821 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5881 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5941 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6001 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6061 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6121 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6181 ctgtggataa ccgtattacc gccatgcat
//
