LOCUS       sequence-177529-        5811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization by Super-Resolution microscopy of beta-Actin fused to 
            mOrange2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  TITLE     Unpublished Henriques Super-Resolution Microscopy Plasmids
REFERENCE   2  (bases 1 to 5811)
  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: RHPMP133 (mOrange2-beta Actin)
FEATURES             Location/Qualifiers
     source          1..5811
                     /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      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             610..1317
                     /codon_start=1
                     /product="enhanced orange to far-red photoswitchable 
                     monomeric fluorescent protein (Subach et al., 2012)"
                     /label=PSmOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGTVNGHEFEIEGEGEGHPYEG
                     FQTAKLKVTKGGPLPFAWDILSPLITYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIRMRLKLKDGGHYTSEVKTTYKAKKSVLLPGAYIVGIKLDITSHNEDYTIVEQYERS
                     EARHSTGGMDELYK"
     primer_bind     1217..1240
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2599..2720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2636..2655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2690..2709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2727..3182)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2864..2885)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3076..3095
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3209..3313
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3315..3672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3315..3506
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3338..3358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3523..3658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3707..4501
                     /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             3707..4501
                     /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(3761..3780)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4371..4390
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4689..4708)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4733..4780
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5109..5697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5598..5617
                     /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 gctagcggtc
      601 gccgccacca tggtgagcaa gggcgaggag aataatatgg ccatcattaa ggagttcatg
      661 cgcttcaagg tgcacatgga gggcactgtg aacggccacg agttcgagat cgagggcgag
      721 ggcgagggcc acccctacga gggctttcag accgctaagc tgaaggtgac caagggcggc
      781 cccctgccct tcgcctggga catcctgtcc cctctcatca cctacggctc caaggcctac
      841 gtgaagcacc ccgccgacat ccccgactac ttcaagctgt ccttccccga gggcttcaag
      901 tgggagcgcg tgatgaacta cgaggacggc ggcgtggtga ccgtgaccca ggactcctct
      961 ctgcaggacg gcgagttcat ctacaaggtg aagatgcgcg gcaccaactt cccctccgac
     1021 ggccccgtga tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1081 gaggacggcg ccctgaaggg cgagatcagg atgaggctga agctgaagga tggcggccac
     1141 tacacctccg aggtcaagac tacctacaag gccaagaagt ccgtgctgct gcccggcgcc
     1201 tacatcgtcg gcatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1261 cagtacgaac gctccgaggc ccgccactcc accggcggca tggacgagct gtacaagtcc
     1321 ggactagatc tcgatcgagc tatggatgat gatatcgccg cgctcgtcgt cgacaacggc
     1381 tccggcatgt gcaaggccgg cttcgcgggc gacgatgccc cccgggccgt cttcccctcc
     1441 atcgtggggc gccccaggca ccagggcgtg atggtgggca tgggtcagaa ggattcctat
     1501 gtgggcgacg aggcccagag caagagaggc atcctcaccc tgaagtaccc catcgagcac
     1561 ggcatcgtca ccaactggga cgacatggag aaaatctggc accacacctt ctacaatgag
     1621 ctgcgtgtgg ctcccgagga gcaccccgtg ctgctgaccg aggcccccct gaaccccaag
     1681 gccaaccgcg agaagatgac ccagatcatg tttgagacct tcaacacccc agccatgtac
     1741 gttgctatcc aggctgtgct atccctgtac gcctctggcc gtaccactgg catcgtgatg
     1801 gactccggtg acggggtcac ccacactgtg cccatctacg aggggtatgc cctcccccat
     1861 gccatcctgc gtctggacct ggctggccgg gacctgactg actacctcat gaagatcctc
     1921 accgagcgcg gctacagctt caccaccacg gccgagcggg aaatcgtgcg tgacattaag
     1981 gagaagctgt gctacgtcgc cctggacttc gagcaagaga tggccacggc tgcttccagc
     2041 tcctccctgg agaagagcta cgagctgcct gacggccagg tcatcaccat tggcaatgag
     2101 cggttccgct gccctgaggc actcttccag ccttccttcc tgggcatgga gtcctgtggc
     2161 atccacgaaa ctaccttcaa ctccatcatg aagtgtgacg tggacatccg caaagacctg
     2221 tacgccaaca cagtgctgtc tggcggcacc accatgtacc ctggcattgc cgacaggatg
     2281 cagaaggaga tcactgccct ggcacccagc acaatgaaga tcaagatcat tgctcctcct
     2341 gagcgcaagt actccgtgtg gatcggcggc tccatcctgg cctcgctgtc caccttccag
     2401 cagatgtgga tcagcaagca ggagtatgac gagtccggcc cctccatcgt ccaccgcaaa
     2461 tgcttctagg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     2521 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2581 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2701 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2761 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2821 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2881 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2941 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3001 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3061 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3121 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3181 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3301 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3361 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3421 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3601 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3661 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3721 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3781 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3841 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3901 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3961 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4021 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4081 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4141 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4201 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4261 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4321 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4381 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4441 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4501 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4561 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4621 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4681 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4741 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4801 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4861 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4921 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4981 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5041 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5101 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5161 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5221 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5281 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5341 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5401 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5461 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5521 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5581 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5641 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5701 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5761 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
