LOCUS       sequence-218675-        6059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Gap Junctions, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6059)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 6059)
  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: mTagRFP-T2-Cx43-7
FEATURES             Location/Qualifiers
     source          1..6059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2039..2048
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2045..2776
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2901..3022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2938..2957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2992..3011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3029..3484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3166..3187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3378..3397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3511..3615
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3617..3974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3617..3808
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3640..3660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3825..3960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4009..4803
                     /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             4009..4803
                     /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(4063..4082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4673..4692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4991..5010)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5035..5082
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5411..5999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5900..5919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      661 tcagatctcg agctcaagct tcgaattcgg cagcactttt ctttcattgg ggggaaggcg
      721 tgaggaaagt accaaacagc agcagacttt taaactttaa acagacaggt ctgagagcct
      781 gaactctcat ttttcctttg acttcagcct ccaaggagtt ccaccaactt tggcgcgccg
      841 gcttcacttt cattaagtga aagagaggtg cccagacatg ggtgactgga gtgccttggg
      901 gaagcttctg gacaaggtcc aagcctactc caccgctgga gggaaggtgt ggctgtcagt
      961 gctcttcata ttcagaatcc tgctcctggg gacagctgtt gagtcagctt ggggtgatga
     1021 acagtctgcc tttcgctgta acactcaaca acctggctgc gaaaacgtct gctatgacaa
     1081 gtccttcccc atctctcacg tgcgcttctg ggtccttcag atcatattcg tgtctgtgcc
     1141 cacactcctg tacttggccc atgtgttcta tgtgatgagg aaggaagaga agctaaacaa
     1201 gaaagaagag gagctcaaag tggcccagac tgacggggtc aacgtggaga tgcacctgaa
     1261 gcagattgaa atcaagaagt tcaagtacgg gattgaagag cacggcaagg tgaaaatgag
     1321 gggcggcttg ctgagaacct acatcatcag catcctcttc aagtctgtct tcgaggtggc
     1381 cttcctgctc atccagtggt acatctatgg gttcagcttg agcgcggtct acacctgcaa
     1441 gagagatccc tgcccccacc aggtagactg cttcctctca cgtcccacgg agaaaaccat
     1501 cttcatcatc ttcatgctgg tggtgtcctt ggtgtctctc gctttgaaca tcattgagct
     1561 cttctacgtc ttcttcaagg gcgttaagga tcgcgtgaag ggaagaagcg atccttacca
     1621 cgccaccact ggcccactga gcccatcaaa agactgcgga tctccaaaat acgcctactt
     1681 caatggctgc tcctcaccaa cggctccact ctcgcctatg tctcctcctg ggtacaagct
     1741 ggttactggt gacagaaaca attcctcgtg ccgcaattac aacaagcaag ctagcgagca
     1801 aaactgggcg aactacagcg cagagcaaaa tcgcatgggg caggccggaa gcaccatctc
     1861 caactcgcac gcccagccgt tcgatttccc cgacgacaac cagaatgcca aaaaagttgc
     1921 tgctggacat gaactccagc cattagccat cgtggaccaa cgaccttcca gcagagccag
     1981 cagccgcgcc agcagcaggc ctcggcctga tgacctggag attgcggatc caccggtcgc
     2041 caccatggtg tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat
     2101 ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta
     2161 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
     2221 cgacatcctg gctaccagct tcatgtacgg cagcagaacc ttcatcaacc acacccaggg
     2281 catccccgat ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     2341 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
     2401 catctacaac gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa
     2461 gaaaacactc ggctgggagg ccaacaccga gaccctgtac cccgctgacg gcggcctgga
     2521 aggcagaacc gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttcaa
     2581 gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt
     2641 ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga
     2701 ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacaaac ttaatggcat
     2761 ggacgagctg tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg
     2821 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2881 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2941 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3001 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3061 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3121 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3181 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3241 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3301 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3361 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3421 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3481 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3601 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3901 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3961 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4021 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4081 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4141 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4201 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4261 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4321 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4381 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4441 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4501 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4561 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4621 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4681 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4741 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4801 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4861 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4921 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4981 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5041 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5101 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5161 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5221 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5521 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatg
//
