LOCUS       sequence-212993-        5218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus/Centromeres, Excitation: 462, Emission: 492.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  AUTHORS   Ai HW, Olenych SG, Wong P, Davidson MW, Campbell RE
  TITLE     Hue-shifted monomeric variants of Clavularia cyan fluorescent 
            protein: identification of the molecular determinants of color and 
            applications in fluorescence imaging.
  JOURNAL   BMC Biol. 2008 Mar 6;6:13. doi: 10.1186/1741-7007-6-13.
  PUBMED    18325109
REFERENCE   2  (bases 1 to 5218)
  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: "BMC Biol.";
            date: "2008-03-6"; volume: "6:13. doi"; pages: " 
            10.1186/1741-7007-6-13"
COMMENT     Sequence Label: mTFP1-CENPB-N-22
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      653..662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1166..1218
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1232..1942
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    2064..2185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2101..2120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2155..2174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2192..2647)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2329..2350)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2541..2560
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2674..2778
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2780..3137
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2780..2971
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2803..2823)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2988..3123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3172..3966
                     /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             3172..3966
                     /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(3226..3245)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3836..3855
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4154..4173)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4198..4245
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4574..5162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5063..5082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagccaccat
      661 gggccccaag cggcggcagc tgacgttccg ggagaagtcg cggatcatcc aggaggtgga
      721 ggagaacccg gacctacgca agggcgagat cgcgcggcgc ttcaacatcc cgccgtccac
      781 gctgagcacg atcctgaaga acaagcgcgc catcctggcg tcggagcgca agtacggggt
      841 ggcctccacc tgccgcaaga ccaacaagct gtctccctac gacaagctcg agggcttgct
      901 catcgcctgg ttccagcaga tccgcgccgc cggcctgccg gtcaagggca tcatcctcaa
      961 ggagaaggcg ctgcgcatag ccgaggagct gggcatggac gacttcaccg cctccaacgg
     1021 ctggctggac cgcttccgcc ggcgccacgg cgtggtgtcc tgcagcggcg tggcccgcgc
     1081 ccgcgcgcga aacgctgccc cccgcacccc ggcggcgcct gccagtccgg ccgcggtgcc
     1141 ctcggagggc agtggcggga gcactctcga gctcaagctt cgaattctgc agtcgacggt
     1201 accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg agaccacaat
     1261 gggcgtaatc aagcccgaca tgaagatcaa gctgaagatg gagggcaacg tgaatggcca
     1321 cgccttcgtg atcgagggcg agggcgaggg caagccctac gacggcacca acaccatcaa
     1381 cctggaggtg aaggagggag cccccctgcc cttctcctac gacattctga ccaccgcgtt
     1441 cgcctacggc aacagggcct tcaccaagta ccccgacgac atccccaact acttcaagca
     1501 gtccttcccc gagggctact cttgggagcg caccatgacc ttcgaggaca agggcatcgt
     1561 gaaggtgaag tccgacatct ccatggagga ggactccttc atctacgaga tacacctcaa
     1621 gggcgagaac ttccccccca acggccccgt gatgcagaag aagaccaccg gctgggacgc
     1681 ctccaccgag aggatgtacg tgcgcgacgg cgtgctgaag ggcgacgtca agcacaagct
     1741 gctgctggag ggcggcggcc accaccgcgt tgacttcaag accatctaca gggccaagaa
     1801 ggcggtgaag ctgcccgact atcactttgt ggaccaccgc atcgagatcc tgaaccacga
     1861 caaggactac aacaaggtga ccgtttacga gagcgccgtg gcccgcaact ccaccgacgg
     1921 catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc cataccacat
     1981 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2041 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2101 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2161 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     2221 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2281 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2341 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2401 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2461 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2521 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2581 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2641 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2761 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2821 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2881 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2941 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3001 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3061 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3121 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3181 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3241 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3301 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3361 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3421 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3481 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3541 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3601 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3661 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3721 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3781 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3841 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3901 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3961 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4021 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4081 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4141 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4201 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4261 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4321 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4381 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4441 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4501 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4561 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4621 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4681 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4741 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4801 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4861 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4921 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4981 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5041 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5101 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5161 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctca
//
