LOCUS       sequence-138503-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4733)
  AUTHORS   Rizzo MA, Springer GH, Granada B, Piston DW
  TITLE     An improved cyan fluorescent protein variant useful for FRET.
  JOURNAL   Nat Biotechnol. 2004 Apr . 22(4):445-9.
  PUBMED    14990965
REFERENCE   2  (bases 1 to 4733)
  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: "Nat 
            Biotechnol. 2004 Apr . 22(4):445-9."
COMMENT     Sequence Label: Cerulean-N1
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    546..626
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(634..636,637..639,640..1353)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(679..700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(940..959)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1287..1308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1476..1597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1513..1532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1567..1586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1604..2059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1741..1762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1953..1972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2086..2190
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2192..2549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2192..2383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2215..2235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2400..2535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2584..3378
                     /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             2584..3378
                     /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(2638..2657)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3248..3267
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3566..3585)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3610..3657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3986..4574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4475..4494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      601 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
      661 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      721 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      781 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
      841 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      901 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      961 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1021 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacgc catcagcgac
     1081 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1141 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1201 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1261 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1321 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     1381 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1441 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1501 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1561 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     1621 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1681 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1741 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1801 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1861 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1921 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1981 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2041 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2101 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2161 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2221 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2281 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2341 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2401 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2461 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2521 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     2581 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2641 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2701 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2761 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2821 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2881 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2941 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3001 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3061 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3121 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3181 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3241 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3301 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3361 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3421 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3481 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3541 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3601 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3661 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3721 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3781 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     3841 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3901 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3961 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4021 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4081 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4141 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4201 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4261 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4321 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4381 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4441 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4501 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4561 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4621 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4681 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag ccc
//
