LOCUS       sequence-420524-        6012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human C16orf59 N-terminally fused to 
            mCerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6012)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6012)
  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"
COMMENT     Sequence Label: pmCer-C16orf59
FEATURES             Location/Qualifiers
     source          1..6012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(624..626,627..629,630..1340)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(669..690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(930..949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1277..1298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2811..2932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2848..2867)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2902..2921
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2939..3394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3076..3097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3288..3307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3421..3525
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3527..3884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3527..3718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3550..3570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3735..3870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3919..4713
                     /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             3919..4713
                     /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(3973..3992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4583..4602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4901..4920)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4945..4992
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5321..5909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5810..5829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      781 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttcg
      841 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1021 aggacggcaa catcctgggg cacaagctgg agtacaacgc catcagcgac aacgtctata
     1081 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1201 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1321 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg aattccttca
     1381 tgctgccggc gggctgctcg cgccggctgg tggccgagct gcagggcgcc ctggacgcct
     1441 gcgcacagcg acaattgcaa ttggagcaga gcctgcgcgt ttgccgtcgg ctgctgcatg
     1501 cctgggaacc aactgggacc cgggctttga agccacctcc agggccagaa actaatggag
     1561 aggaccccct tccagcatgc acacccagtc cacaagacct caaagagttg gagtttctga
     1621 cccaggcact ggagaaggct gtacgagttc gaagaggcat cactaaggcc ggagagagag
     1681 acaaggcccc cagcctgaaa tctaggtcca ttgtcacctc ttctggcacg acagcctccg
     1741 ccccaccgca ttccccaggc caagctggtg gccatgcttc agacacgaga cccaccaagg
     1801 gcctccgcca gaccacggtg cctgccaagg gccaccctga gcgccggctg ctgtcagtgg
     1861 gggatgggac ccgtgttggg atgggagccc gaacccccag gcctggggcg ggcctcaggg
     1921 accagcaaat ggccccatcc gctgctcctc aggccccaga agccttcaca ctcaaggaga
     1981 aggggcacct gctgcggctg cctgcggcat tcaggaaagc agcttcccag aactcgagcc
     2041 tgtgggccca gctcagttcc acacagacca gtgattccac ggatgccgcc gctgccaaaa
     2101 cccagttcct ccagaacatg cagacagctt caggcgggcc ccagcccagg ctcagtgctg
     2161 tggaggtgga ggcggaggcg gggcgcctgc ggaaggcctg ctcgctgctg agactgcgca
     2221 tgagggagga gctctcggca gcccccatgg actggatgca ggagtaccgc tgcctgctca
     2281 cgctggaggg gctgcaggcc atggtgggcc agtgtctgca caggctgcag gagctgcgtg
     2341 cagcggtggc ggaacagcca ccaagaccat gtcctgtggg gaggcccccc ggagcctcgc
     2401 cgtcctgtgg gggtagagcg gagcctgcat ggagccccca gctgcttgtc tactccagca
     2461 cccaggagct gcagaccctg gcggccctca agctgcgagt ggctgtgctg gaccagcaga
     2521 tccacttgga aaaggtcctg atggctgaac tcctccccct ggtaagcgct gcacagccgc
     2581 aggggccgcc ctggctggcc ctgtgccggg ctgtgcacag cctgctctgc gagggaggag
     2641 cacgtgtcct taccatcctg cgggatgaac ctgcagtctg aggatccacc ggatctagat
     2701 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2761 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2821 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2881 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     2941 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3001 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3061 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3121 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3181 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3241 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3301 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3361 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3481 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3541 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3841 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3901 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3961 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4021 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4081 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4141 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4201 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4261 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4321 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4381 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4441 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4501 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4561 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4621 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4681 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4741 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4801 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4861 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4921 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4981 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5041 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5101 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5161 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5221 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5281 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5341 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5401 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5461 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtatcaccgc
     5521 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5581 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5641 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5701 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5761 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5821 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5881 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5941 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6001 ataaccgtat ta
//
