LOCUS       sequence-182207-        5377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Golgi/ER, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5377)
  TITLE     Davidson Cyan Fluorescent Proteins
REFERENCE   2  (bases 1 to 5377)
  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: ECFP-ELP1-25
FEATURES             Location/Qualifiers
     source          1..5377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      540..549
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     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"
     polyA_signal    2105..2226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2142..2161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2196..2215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2233..2688)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2370..2391)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2582..2601
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2715..2819
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2821..3178
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2821..3012
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2844..2864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3029..3164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3213..4007
                     /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             3213..4007
                     /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(3267..3286)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3877..3896
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4195..4214)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4239..4286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4615..5203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5104..5123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgccg
      541 ccaccatggc catgaacatt ttccggctga ctggggacct gtcccacctg gcggccatcg
      601 tcatcctgct gctgaagatc tggaagacgc gctcctgcgc cggtatttct gggaaaagcc
      661 agcttctgtt tgcactggtc ttcacaactc gttacctgga tctttttact tcatttattt
      721 cattgtataa cacatctatg aaggttatct accttgcctg ctcctatgcc acagtgtacc
      781 tgatctacct gaaatttaag gcaacctacg atggaaatca tgataccttc cgagtggagt
      841 ttctggtggt ccctgtggga ggcctctcat ttttagttaa tcacgatttc tctcctcttg
      901 agatcctctg gaccttctcc atctacctgg agtccgtggc tatccttccg cagctgttta
      961 tgatcagcaa gactggggag gccgagacca tcaccaccca ctacctgttc ttcctgggcc
     1021 tctatcgtgc tttgtatctt gtcaactgga tctggcgctt ctactttgag ggcttctttg
     1081 acctcattgc tgtggtggcc ggcgtagtcc agaccatcct atactgtgac ttcttctact
     1141 tgtacattac aaaagtactc aagggaaaga agctcagttt gccagcaggc ggtggggggg
     1201 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     1261 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1321 acggcgacgt aaacggccac aggttcagcg tgtccggcga gggcgagggc gatgccacct
     1381 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1441 ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc gaccacatga
     1501 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgtaccatct
     1561 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1621 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1681 acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac aagcagaaga
     1741 acggcatcaa ggcccacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1801 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1861 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1921 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1981 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2041 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2101 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2161 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2221 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2281 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2341 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2401 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2461 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2521 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2581 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2641 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2701 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2761 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     2821 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2881 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3061 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3121 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3181 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3241 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3301 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3361 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3421 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3481 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3541 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3601 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3661 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3721 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3781 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3841 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3901 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3961 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4021 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4081 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4141 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4201 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4261 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4321 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4381 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4441 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     4501 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4561 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4621 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4681 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4741 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4801 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4861 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4921 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4981 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5041 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5101 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5161 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5221 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5281 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     5341 cggggtcatt agttcatagc ccatatatgg agttccg
//
