LOCUS       sequence-202952-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a circularly permuted mCerulean3 for FRET sensor optimization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4749)
  AUTHORS   Markwardt ML, Seckinger KM, Rizzo MA
  TITLE     Regulation of Glucokinase by Intracellular Calcium Levels in 
            Pancreatic beta Cells.
  JOURNAL   J Biol Chem. 2016 Feb 5;291(6):3000-9. doi: 10.1074/jbc.M115.692160.
            Epub 2015 Dec 23.
  PUBMED    26698632
REFERENCE   2  (bases 1 to 4749)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M115.692160"; journalName: "J Biol Chem"; date: 
            "2016-02-5- 5"; volume: "291"; issue: "6"; pages: "3000-9"
COMMENT     Sequence Label: cp173-mCer3-C1
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     571..591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     802..823
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(926..947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1187..1206)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1400..1465
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1589..1710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1626..1645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1680..1699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1717..2172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1854..1875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2066..2085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2199..2303
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2305..2662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2305..2496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2328..2348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2513..2648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2697..3491
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2697..3491
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2751..2770)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3361..3380
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3679..3698)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3723..3770
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4099..4687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4588..4607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      661 caccatggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
      721 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
      781 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
      841 tctcggcatg gacgagctgt acaagggtgg cagcggtggc atggtgagca agggcgagga
      901 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      961 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1021 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagctg
     1081 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1141 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1201 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1261 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat
     1321 ccacggcaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcgg
     1381 cctcaactgc aacatcgagt ccggactcag atctcgagct caagcttcga attctgcagt
     1441 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     1501 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1561 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1621 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1681 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     1741 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1801 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1861 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1921 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1981 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2041 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2101 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2161 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2221 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2281 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2341 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2401 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2461 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2521 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2581 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2641 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2701 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2761 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2821 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2881 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2941 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3001 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3061 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3121 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3181 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3241 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3301 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3361 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3421 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3481 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3541 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3661 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3721 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3781 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3841 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3901 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3961 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4021 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4081 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4141 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4201 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4261 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4321 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4381 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4441 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4501 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4561 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4621 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4681 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4741 tcacatgtt
//
