LOCUS       sequence-419424-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of the N-terminal amino acids 1-87 of human 
            C17orf53 C-terminally fused to mCerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6415)
  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-C17orf53_1-87-mCer
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(68..87)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        259..638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     793..813
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     887..906
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        887..905
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1257..1266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1263..1265,1266..1268,1269..1982)
                     /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(1308..1329)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1569..1588)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1916..1937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1992..2010)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1993..2010)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2030..2047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2036..2260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2306..2734
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2393..2412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2603..2624
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2743..2763)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2748..3077
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2928..3063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2990..3009
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3144..3938
                     /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(3198..3217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3808..3827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4112..4233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4149..4168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4203..4222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4282..4298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4282..4298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4295..4317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4306..4322
                     /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(4330..4336,4337..4354,4355..4360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4375..4396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4513..4530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4684..5272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4764..4783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5443..6234,6235..6303))
                     /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     6066..6085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6304..6408)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
       61 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      121 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      181 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      241 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      301 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      361 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      421 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      481 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      541 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      601 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      661 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      721 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      781 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      841 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      901 atagggagac ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct
      961 ggaattctat ggcgtgcagt ttgcagaagc tgtttgctgt ggaagaggag tttgaagatg
     1021 aggatttctt gtctgctgtg gaggatgcag agaaccggtt tactggctca ctgcctgtga
     1081 atgctgggcg cctgagacct gtctcttcta ggccacagga gactgtgcag gcacagtcct
     1141 ccaggctgct gctgttacac cccactgctc cctcagaggc tttgggcctg ccagacttgg
     1201 acctctgcct ccctgcctcc agcacgcccc tcgagcatgc atctagaaga tccatcgcca
     1261 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1321 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1381 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1441 ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     1501 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1561 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1621 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1681 acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac aagcagaaga
     1741 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1801 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1861 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1921 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1981 aagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     2041 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2101 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2161 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2221 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2281 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2341 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2401 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2461 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2521 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2581 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2641 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2701 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2821 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2881 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2941 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3001 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3061 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3121 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3301 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3661 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4081 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4141 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4201 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4261 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4321 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4381 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4441 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4501 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4561 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4621 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4681 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4801 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4861 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4921 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4981 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5041 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5101 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5161 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5221 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca cattt
//
