LOCUS       sequence-368052-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LSSmGFP fluorescent protein (ex/em = 400/510 nm): cytosolic 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
REFERENCE   2  (bases 1 to 6129)
  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"; journalName: "Nat 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: pcDNA3.1-LSSmGFP
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     746..765
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        746..764
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(875..896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1565..1583)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1566..1583)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1603..1620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1609..1833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1879..2307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1966..1985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2176..2197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2316..2336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2321..2650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2501..2636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2563..2582
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2717..3511
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2771..2790)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3381..3400
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3722..3741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3776..3795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3855..3871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3855..3871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3868..3890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3879..3895
                     /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(3903..3909,3910..3927,3928..3933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3948..3969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4086..4103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4257..4845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4337..4356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5016..5807,5808..5876))
                     /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     5639..5658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5877..5981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6056..6075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
       61 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      721 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt
      781 accgagctcg gatccactag taacggccgc cagtgtgctg gaattcggca tggtgagcaa
      841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      901 cggccacaag ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaaactgtc
      961 cctgaagctc atctccacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1021 cttaagctac ggccagatgg tgttcgcccg ctaccctgac aacatgaagc agcacgactt
     1081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatctctt tcgaggacga
     1141 cggttactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1201 cgtgctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca acctggagta
     1261 caacttcaac agccacaacg tctatatcac ggccgacaag cagaagaacg gcatcaaggc
     1321 taacttcaag atccgccaca acgttgagga cggcggcgtg cagctcgccg accactacca
     1381 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcat
     1441 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgaaggagag
     1501 ggtgaccgcc gccgggatta cacatgacat gaacgagctg tacaagtaat ctagagggcc
     1561 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     1621 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1681 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1741 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1801 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1861 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1921 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1981 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2041 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2101 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2161 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2221 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2281 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2401 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2461 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2521 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2581 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2641 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     2701 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2761 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2821 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2881 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2941 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3001 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3061 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3121 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3181 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3241 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3301 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3361 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3421 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3481 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3541 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3601 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3661 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3781 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4081 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4921 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4981 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5041 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5101 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5161 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5221 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5281 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5341 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5401 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5461 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5521 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5581 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5641 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5701 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5761 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5821 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5941 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6001 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6061 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6121 ctgagtagt
//
