LOCUS       sequence-194902-        6182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein amFP486/K68M under a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6182)
  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 6182)
  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: pThom77.6
FEATURES             Location/Qualifiers
     source          1..6182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    19..243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      289..717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(376..395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     586..607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(726..746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        731..1060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      911..1046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     973..992
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2262..2383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2299..2318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2353..2372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2432..2448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2432..2448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2445..2467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2456..2472
                     /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(2480..2486,2487..2504,2505..2510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2525..2546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2663..2680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2834..3422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2914..2933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3593..4384,4385..4453))
                     /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     4216..4235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4454..4558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4633..4652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4824..5203
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5204..5407
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5358..5378
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5452..5471
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5452..5470
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5494..5503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cttctaacgc gccctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
       61 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
      121 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
      181 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
      241 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
      301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      361 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      421 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      481 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      601 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
      661 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
      721 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
      781 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
      841 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
      901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
      961 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     1021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     1081 atccattttc ggatctgatc agcacgtgat gaaaaagcct gaactcaccg cgacgtctgt
     1141 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     1201 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     1261 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     1321 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     1381 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     1441 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     1501 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     1561 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     1621 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     1681 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     1741 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     1801 catcttcttc tggaggccgt ggttggcttg tagagcagca gacgcgctac ttcgagcgga
     1861 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     1921 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     1981 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     2041 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     2101 gccccagcac tcgtccgagg gcaaaggaat agcacgtgct acgagatttc gattccaccg
     2161 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     2221 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     2281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     2401 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2461 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2521 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4561 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     4621 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     4681 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     4741 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     4801 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     4861 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4921 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4981 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5041 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5101 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5161 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     5221 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5281 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     5341 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     5401 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     5461 tcactatagg gagacccaag ctggctagcg tttgccacca tggccctgag caacaagttc
     5521 atcggcgacg acatgaagat gacctaccac atggacggct gcgtgaacgg ccactacttc
     5581 accgtgaagg gcgagggcaa cggcaagccc tacgagggca cccagaccag caccttcaag
     5641 gtgaccatgg ccaacggcgg ccccctggcc ttcagcttcg acatcctgag caccgtgttc
     5701 atgtacggca accggtgctt caccgcctac cccaccagca tgcccgacta cttcaagcag
     5761 gccttccccg acggcatgag ctacgagcgg accttcacct acgaggacgg cggcgtggcc
     5821 accgccagct gggagatcag cctgaagggc aactgcttcg agcacaagag caccttccac
     5881 ggcgtgaact tccccgccga cggccccgtg atggccaaga agaccaccgg ctgggacccc
     5941 agcttcgaga agatgaccgt gtgcgacggc atcctgaagg gcgacgtgac cgccttcctg
     6001 atgctgcagg gcggcggcaa ctaccggtgc cagttccaca ccagctacaa gaccaagaag
     6061 cccgtgacca tgccccccaa ccacgtggtg gagcaccgga tcgcccggac cgacctggac
     6121 aagggcggca acagcgtgca gctgaccgag cacgccgtgg cccacatcac cagcgtggtg
     6181 cc
//
