LOCUS       sequence-321577-        6180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses both eGFP and chicken ovalbumin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6180)
  TITLE     Expression plasmid for production of murine IFN gamma
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6180)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: eGFP-ova-C1
FEATURES             Location/Qualifiers
     source          1..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1450..2610
                     /codon_start=1
                     /product="chicken ovalbumin"
                     /label=ovalbumin
                     /translation="MGSIGAASMEFCFDVFKELKVHHANENIFYCPIAIMSALAMVYLG
                     AKDSTRTQINKVVRFDKLPGFGDSIEAQCGTSVNVHSSLRDILNQITKPNDVYSFSLAS
                     RLYAEERYPILPEYLQCVKELYRGGLEPINFQTAADQARELINSWVESQTNGIIRNVLQ
                     PSSVDSQTAMVLVNAIVFKGLWEKAFKDEDTQAMPFRVTEQESKPVQMMYQIGLFRVAS
                     MASEKMKILELPFASGTMSMLVLLPDEVSGLEQLESIINFEKLTEWTSSNVMEERKIKV
                     YLPRMKMEEKYNLTSVLMAMGITDVFSSSANLSGISSAESLKISQAVHAAHAEINEAGR
                     EVVGSAEAGVDAASVSEEFRADHPFLFCIKHIATNAVLFFGRCVSP"
     polyA_signal    2968..3089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3005..3024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3059..3078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3096..3551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3233..3254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3445..3464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3578..3682
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3684..4041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3684..3875
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3707..3727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3892..4027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4076..4870
                     /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             4076..4870
                     /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(4130..4149)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4740..4759
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5058..5077)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5102..5149
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5478..6066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5967..5986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcaga atatcagaaa
     1381 cagtttttaa ttgcgttctc gcagccggta ccgagctcga attcctcgaa agacaactca
     1441 gagttcacca tgggctccat cggtgcagca agcatggaat tttgttttga tgtattcaag
     1501 gagctcaaag tccaccatgc caatgagaac atcttctact gccccattgc catcatgtca
     1561 gctctagcca tggtatacct gggtgcaaaa gacagcacca ggacacaaat aaataaggtt
     1621 gttcgctttg ataaacttcc aggattcgga gacagtattg aagctcagtg tggcacatct
     1681 gtaaacgttc actcttcact tagagacatc ctcaaccaaa tcaccaaacc aaatgatgtt
     1741 tattcgttca gccttgccag tagactttat gctgaagaga gatacccaat cctgccagaa
     1801 tacttgcagt gtgtgaagga actgtataga ggaggcttgg aacctatcaa ctttcaaaca
     1861 gctgcagatc aagccagaga gctcatcaat tcctgggtag aaagtcagac aaatggaatt
     1921 atcagaaatg tccttcagcc aagctccgtg gattctcaaa ctgcaatggt tctggttaat
     1981 gccattgtct tcaaaggact gtgggagaaa gcatttaagg atgaagacac acaagcaatg
     2041 cctttcagag tgactgagca agaaagcaaa cctgtgcaga tgatgtacca gattggttta
     2101 tttagagtgg catcaatggc ttctgagaaa atgaagatcc tggagcttcc atttgccagt
     2161 gggacaatga gcatgttggt gctgttgcct gatgaagtct caggccttga gcagcttgag
     2221 agtataatca actttgaaaa actgactgaa tggaccagtt ctaatgttat ggaagagagg
     2281 aagatcaaag tgtacttacc tcgcatgaag atggaggaaa aatacaacct cacatctgtc
     2341 ttaatggcta tgggcattac tgacgtgttt agctcttcag ccaatctgtc tggcatctcc
     2401 tcagcagaga gcctgaagat atctcaagct gtccatgcag cacatgcaga aatcaatgaa
     2461 gcaggcagag aggtggtagg gtcagcagag gctggagtgg atgctgcaag cgtctctgaa
     2521 gaatttaggg ctgaccatcc attcctcttc tgtatcaagc acatcgcaac caacgccgtt
     2581 ctcttctttg gcagatgtgt ttccccttaa aaagaagaaa gctgaaaaac tctgtccctt
     2641 ccaacaagac ccagagcact gtagtatcag gggtaaaatg aaaagtatgt tatctgctgc
     2701 atccagactt cataaaagct ggagcttaat ctagaaaaaa aatcagaaag aaattacact
     2761 gtgagaacag gtgcaattca cttttccttt acacagagta atactggtaa ctcatggatg
     2821 aaggcttaag agtgataggg atccaccgga tctagataac tgatcataat cagccatacc
     2881 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2941 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3001 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3061 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     3121 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3181 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3241 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3301 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3361 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3421 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3481 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3541 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3601 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3661 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3721 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3841 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3901 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3961 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4021 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4081 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4141 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4201 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4261 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4321 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4381 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4441 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4501 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4561 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4621 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4681 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4741 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4801 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4861 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4921 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4981 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5041 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5101 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5161 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5221 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5281 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5341 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5401 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5461 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5521 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5581 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5641 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5701 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5761 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5821 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5881 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5941 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6001 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6061 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6121 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
