LOCUS       sequence-191738-        6200 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 6200)
  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 6200)
  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..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..23)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12..236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      282..710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(369..388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     579..600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(719..739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        724..1053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      904..1039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     966..985
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1102..2127
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1102..2127
                     /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    2257..2378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2294..2313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2348..2367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2427..2443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2427..2443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2440..2462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2451..2467
                     /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(2475..2481,2482..2499,2500..2505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2520..2541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2658..2675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2829..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2909..2928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3585..4376,4377..4445))
                     /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     4208..4227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4446..4509)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4644..4663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4835..5214
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5215..5418
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5369..5389
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5463..5482
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5463..5481
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5505..5514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgcgccctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
       61 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
      121 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
      181 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
      241 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
      301 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      361 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      421 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      481 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      541 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      601 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      661 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
      721 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      781 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      841 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      901 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      961 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     1021 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     1081 ttcggatctg atcagcacgt gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     1141 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     1201 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     1261 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     1321 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     1381 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     1441 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     1501 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     1561 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     1621 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     1681 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     1741 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     1801 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     1861 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     1921 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     1981 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     2041 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     2101 agcactcgtc cgagggcaaa ggaatagcac gtgctacgag atttcgattc caccgccgcc
     2161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     2221 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     2281 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2341 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     2401 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2461 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2521 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2581 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2641 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2701 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2761 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2821 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2881 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2941 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3001 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3061 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3121 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3181 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3241 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3301 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3361 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4501 ggatacatat ttgttttcta aatacattca aatatgtatc cgctcatata aacaaatagg
     4561 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     4621 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     4681 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     4741 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     4801 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     4861 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4921 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4981 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5041 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5101 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5161 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5221 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     5281 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     5341 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     5401 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     5461 attaatacga ctcactatag ggagacccaa gctggctagc gtttgccacc atggccctga
     5521 gcaacaagtt catcggcgac gacatgaaga tgacctacca catggacggc tgcgtgaacg
     5581 gccactactt caccgtgaag ggcgagggca acggcaagcc ctacgagggc acccagacca
     5641 gcaccttcaa ggtgaccatg gccaacggcg gccccctggc cttcagcttc gacatcctga
     5701 gcaccgtgtt catgtacggc aaccggtgct tcaccgccta ccccaccagc atgcccgact
     5761 acttcaagca ggccttcccc gacggcatga gctacgagcg gaccttcacc tacgaggacg
     5821 gcggcgtggc caccgccagc tgggagatca gcctgaaggg caactgcttc gagcacaaga
     5881 gcaccttcca cggcgtgaac ttccccgccg acggccccgt gatggccaag aagaccaccg
     5941 gctgggaccc cagcttcgag aagatgaccg tgtgcgacgg catcctgaag ggcgacgtga
     6001 ccgccttcct gatgctgcag ggcggcggca actaccggtg ccagttccac accagctaca
     6061 agaccaagaa gcccgtgacc atgcccccca accacgtggt ggagcaccgg atcgcccgga
     6121 ccgacctgga caagggcggc aacagcgtgc agctgaccga gcacgccgtg gcccacatca
     6181 ccagcgtggt gcccttctaa
//
