LOCUS       sequence-36039-e        5210 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5210)
  AUTHORS   Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV
  TITLE     A photoswitchable orange-to-far-red fluorescent protein, PSmOrange.
  JOURNAL   Nat Methods. 2011 Jul 31;8(9):771-7. doi: 10.1038/nmeth.1664.
  PUBMED    21804536
REFERENCE   2  (bases 1 to 5210)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-07-31- 31"; volume: "8"; 
            issue: "9"; pages: "771-7"
COMMENT     Sequence Label: pMyosin-PSmOrange
FEATURES             Location/Qualifiers
     source          1..5210
                     /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      1159..1168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1165..1875
                     /codon_start=1
                     /product="orange to far-red photoswitchable variant of the 
                     monomeric fluorescent protein mOrange (Subach et al., 
                     2011)"
                     /label=PSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPLFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIRMRLKLKDGGHYTSEVKTTYKAKKSVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1772..1795
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1998..2119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2035..2054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2089..2108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2126..2581)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2263..2284)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2475..2494
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2608..2712
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2714..3071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2714..2905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2737..2757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2922..3057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3106..3900
                     /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             3106..3900
                     /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(3160..3179)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3770..3789
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4088..4107)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4132..4179
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4508..5096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4997..5016
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttatg tcgagcaaga gagccaaggc caagaccacc
      661 aagaagaggc cccagagggc tacgtccaat gtcttcgcca tgtttgacca gtcccagatc
      721 caggagttta aggaggcctt caacatgatt gatcagaacc gcgatggctt cattgataag
      781 gaggacctgc acgacatgtt ggcctctctg gggaagaacc ccacagacga gtacctggag
      841 ggcatgatga acgaggcgcc agggcccatc aacttcacca tgttcctcac aatgtttggg
      901 gagaagctga acggcacaga ccccgaggat gtgatccgca atgcctttgc ctgctttgat
      961 gaggaggcct caggcttcat ccacgaggac cacctgaggg agctgctcac caccatgggc
     1021 gaccgattca cggatgagga ggtggacgag atgtaccgcg aggcacccat tgataagaag
     1081 ggcaacttca actacgtgga gttcactcgc atcctcaaac acggcgccaa ggacaaggac
     1141 gacctggatc caccggtcgc caccatggtg agcaagggcg aggagaataa tatggccatc
     1201 atcaaggagt tcatgcgctt caaggtgcgc atggagggca ccgtgaacgg ccacgagttc
     1261 gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgc taagctgaag
     1321 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctct cttcacctac
     1381 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttcaa gctgtccttc
     1441 cccgagggct tcaagtggga gcgcgtgatg aactacgagg acggcggcgt ggtgaccgtg
     1501 acccaggact cctcactgca ggacggcgag ttcatctaca aggtgaagat gcgcggcacc
     1561 aacttcccct ccgacggccc cgtgatgcag aagaagacca tgggctggga ggcctcctcc
     1621 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaggatgag gctgaagctg
     1681 aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa gaagtccgtg
     1741 cagctgcccg gcgcctacat cgtcggcatc aagctggaca tcacctccca caacgaggac
     1801 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1861 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1921 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1981 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2101 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2161 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2221 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2281 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2341 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2401 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2461 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2521 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2701 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2821 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2941 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3001 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3061 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3121 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3181 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3241 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3301 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3361 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3421 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3481 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3541 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3601 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3661 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3721 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3781 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3841 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3901 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3961 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4021 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4081 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4141 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4201 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4261 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4321 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4621 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
