LOCUS       sequence-420479-        7457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human C16orf59 C-terminally fused to 
            mCerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 7457)
  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: pcDNA3-C16orf59-mCer
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2277..2286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2283..2285,2286..2288,2289..3002)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2328..2349)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2589..2608)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2936..2957
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3012..3030)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3013..3030)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3050..3067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3056..3280
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3326..3754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3413..3432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3623..3644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3763..3783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3768..4097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3948..4083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4010..4029
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4164..4958
                     /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(4218..4237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4828..4847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5132..5253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5169..5188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5223..5242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5302..5318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5302..5318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5315..5337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5326..5342
                     /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(join(5350..5356,5357..5374,5375..5380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5395..5416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5533..5550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5784..5803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6463..7254,7255..7323))
                     /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     7086..7105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7324..7428)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      901 ccgagctcgg atccttcatg ctgccggcgg gctgctcgcg ccggctggtg gccgagctgc
      961 agggcgccct ggacgcctgc gcacagcgac aattgcaatt ggagcagagc ctgcgcgttt
     1021 gccgtcggct gctgcatgcc tgggaaccaa ctgggacccg ggctttgaag ccacctccag
     1081 ggccagaaac taatggagag gacccccttc cagcatgcac acccagtcca caagacctca
     1141 aagagttgga gtttctgacc caggcactgg agaaggctgt acgagttcga agaggcatca
     1201 ctaaggccgg agagagagac aaggccccca gcctgaaatc taggtccatt gtcacctctt
     1261 ctggcacgac agcctccgcc ccaccgcatt ccccaggcca agctggtggc catgcttcag
     1321 acacgagacc caccaagggc ctccgccaga ccacggtgcc tgccaagggc caccctgagc
     1381 gccggctgct gtcagtgggg gatgggaccc gtgttgggat gggagcccga acccccaggc
     1441 ctggggcggg cctcagggac cagcaaatgg ccccatccgc tgctcctcag gccccagaag
     1501 ccttcacact caaggagaag gggcacctgc tgcggctgcc tgcggcattc aggaaagcag
     1561 cttcccagaa ctcgagcctg tgggcccagc tcagttccac acagaccagt gattccacgg
     1621 atgccgccgc tgccaaaacc cagttcctcc agaacatgca gacagcttca ggcgggcccc
     1681 agcccaggct cagtgctgtg gaggtggagg cggaggcggg gcgcctgcgg aaggcctgct
     1741 cgctgctgag actgcgcatg agggaggagc tctcggcagc ccccatggac tggatgcagg
     1801 agtaccgctg cctgctcacg ctggaggggc tgcaggccat ggtgggccag tgtctgcaca
     1861 ggctgcagga gctgcgtgca gcggtggcgg aacagccacc aagaccatgt cctgtgggga
     1921 ggccccccgg agcctcgccg tcctgtgggg gtagagcgga gcctgcatgg agcccccagc
     1981 tgcttgtcta ctccagcacc caggagctgc agaccctggc ggccctcaag ctgcgagtgg
     2041 ctgtgctgga ccagcagatc cacttggaaa aggtcctgat ggctgaactc ctccccctgg
     2101 taagcgctgc acagccgcag gggccgccct ggctggccct gtgccgggct gtgcacagcc
     2161 tgctctgcga gggaggagca cgtgtcctta ccatcctgcg ggatgaacct gcagtcgaat
     2221 tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaaga tccatcgcca
     2281 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2341 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2401 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2461 ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     2521 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2581 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2641 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2701 acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac aagcagaaga
     2761 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2821 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2881 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2941 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3001 aagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3241 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3301 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3721 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4021 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4081 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4141 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4201 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4261 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4321 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4381 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4441 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4501 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4561 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4621 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4681 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4741 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4801 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4861 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4921 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7441 aaaagtgcca cctgacg
//
