LOCUS       sequence-241190-        5673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5673)
  TITLE     Davidson Yellow fluorescent proteins
REFERENCE   2  (bases 1 to 5673)
  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: EYFP-Annexin-12
FEATURES             Location/Qualifiers
     source          1..5673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(645..647,648..650,651..1361)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(690..711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(951..970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1298..1319
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2493..2614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2530..2549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2584..2603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2621..3076)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2758..2779)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2970..2989
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3103..3207
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3209..3566
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3209..3400
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3232..3252)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3417..3552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3601..4395
                     /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             3601..4395
                     /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(3655..3674)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4265..4284
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4583..4602)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4627..4674
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5003..5591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5492..5511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
      661 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      721 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      781 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
      841 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      901 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      961 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1021 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1081 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1141 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1201 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1261 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1321 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
     1381 ctcaagcttc gaattctatg gccatggcaa ccaaaggagg tactgtcaaa gctgcttcag
     1441 gattcaatgc catggaagat gcccagaccc tgaggaaggc catgaaaggg ctcggcaccg
     1501 atgaagacgc cattattagc gtccttgcct accgcaacac cgcccagcgc caggagatca
     1561 ggacagccta caagagcacc atcggcaggg acttgataga cgacctgaag tcagaactga
     1621 gtggcaactt cgagcaggtg attgtgggga tgatgacgcc cacggtgctg tatgacgtgc
     1681 aagagctgcg aagggccatg aagggagccg gcactgatga gggctgccta attgagatcc
     1741 tggcctcccg gacccctgag gagatccggc gcataagcca aacctaccag cagcaatatg
     1801 gacggagcct tgaagatgac attcgctctg acacatcgtt catgttccag cgagtgctgg
     1861 tgtctctgtc agctggtggg agggatgaag gaaattatct ggacgatgct ctcgtgagac
     1921 aggatgccca ggacctgtat gaggctggag agaagaaatg ggggacagat gaggtgaaat
     1981 ttctaactgt tctctgttcc cggaaccgaa atcacctgtt gcatgtgttt gatgaataca
     2041 aaaggatatc acagaaggat attgaacaga gtattaaatc tgaaacatct ggtagctttg
     2101 aagatgctct gctggctata gtaaagtgca tgaggaacaa atctgcatat tttgctgaaa
     2161 agctctataa atcgatgaag ggcttgggca ccgatgataa caccctcatc agagtgatgg
     2221 tttctcgagc agaaattgac atgttggata tccgggcaca cttcaagaga ctctatggaa
     2281 agtctctgta ctcgttcatc aagggtgaca catctggaga ctacaggaaa gtactgcttg
     2341 ttctctgtgg aggagatgat taaggatcca ccggatctag ataactgatc ataatcagcc
     2401 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2461 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2521 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2581 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2641 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2701 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2761 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2821 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2881 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2941 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3001 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3061 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3121 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3181 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3241 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3301 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3361 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3421 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3481 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3541 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3601 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3661 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3721 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3781 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3841 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3901 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3961 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4021 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4081 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4141 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4201 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4261 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4321 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4381 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4441 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4501 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4561 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4621 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4681 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4741 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4801 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4981 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5041 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5101 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5161 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5221 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5281 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5341 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5401 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5461 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5521 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5581 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5641 ttgctcacat gttctttcct gcgttatccc ctg
//
