LOCUS       sequence-191734-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein Rosmarinus under a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, 
            Hughes TE, Drobizhev M
  TITLE     Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than 
            Enhanced Green Fluorescent Protein under Two-Photon Excitation.
  JOURNAL   J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 
            10.1021/acs.jpclett.7b00960. Epub 2017 May 25.
  PUBMED    28530831
REFERENCE   2  (bases 1 to 6194)
  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"; doi: 
            "10.1021/acs.jpclett.7b00960"; journalName: "J Phys Chem Lett"; 
            date: "2017-06-15- 15"; volume: "8"; issue: "12"; pages: "2548-2554"
COMMENT     Sequence Label: pRSM32.2
FEATURES             Location/Qualifiers
     source          1..6194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(711..728)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    717..941
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      987..1415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1074..1093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1284..1305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1424..1444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1429..1758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1609..1744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1671..1690
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1807..2832
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1807..2832
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    2962..3083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2999..3018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3053..3072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3132..3148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3132..3148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3145..3167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3156..3172
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3180..3186,3187..3204,3205..3210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3225..3246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3363..3380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3534..4119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3614..3633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4290..5081,5082..5150))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4913..4932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5151..5214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5349..5368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5540..5919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5920..6123
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6074..6094
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6168..6187
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6168..6186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agctggctag cgtttgccac catgtcagtg ataaagagcg tcatgaaaat taagctgcat
       61 atggacggca tcgtgaacgg tcacaaattc atgatcaccg gtgaggggga aggcaagcca
      121 tttgagggga ctcataccat tatcctgaag gtgaaggaag gaggaccact gcctttcgca
      181 tatgatatcc tcactacagc ctttcagtac ggcaatcggg tcttcacaaa atatcccaag
      241 gatattcctg actacttcaa acagtcattc ccggagggct acagttggga gcggtccatg
      301 acctttgagg atcagggcgt gtgtaccgtg acctctgaca tcaaactgga aggagactat
      361 ttcacctacg atatcaggtt tcacggagtc aatttccccg ccggcggtcc cgtcatgcag
      421 aaaaagactc tgcggtggga accaagcacc gagaacatgt acgtgcgaga cggcgtgctg
      481 gtaggcgagg tggagagaac tctgctgctg gagggcaata aacatcaccg ctgtaacttc
      541 agaacaactt acaaagcaaa gaaagaagta gtcctgccag aataccactt tgttgaccac
      601 cggatagaga tcttggggca tgacaaggac tataacaatg tggttgtgta tgaaaacgca
      661 gtggctcggc aacaggcctc taccctgcca agcaaggcga agtagcgcgc cctcgactgt
      721 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
      781 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
      841 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
      901 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
      961 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1201 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1321 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1381 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1441 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1501 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1561 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1621 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1681 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1741 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     1801 cacgtgatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     1861 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     1921 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     1981 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     2041 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     2101 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     2161 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     2221 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     2281 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     2341 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     2401 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     2461 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     2521 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     2581 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     2641 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     2701 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     2761 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     2821 gcaaaggaat agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     2881 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     2941 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3001 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3061 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3121 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3181 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3241 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3361 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3421 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     4081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgtt
     5221 ttctaaatac attcaaatat gtatccgctc atataaacaa ataggggttc cgcgcacatt
     5281 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5341 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     5401 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     5461 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     5521 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     5581 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5641 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5701 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5761 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5821 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5881 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5941 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     6001 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     6061 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     6121 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     6181 tatagggaga ccca
//
