LOCUS       sequence-392993-        4811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid IVT template containing a humanized Monster Green 
            Fluorescent Protein (hMGFP) CDS followed by a template-encoded 
            poly(A) tail. Linearization at an Esp3i site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4811)
  AUTHORS   Aditham A, Shi H, Guo J, Zeng H, Zhou Y, Wade SD, Huang J, Liu J, 
            Wang X
  TITLE     Chemically Modified mocRNAs for Highly Efficient Protein Expression 
            in Mammalian Cells.
  JOURNAL   ACS Chem Biol. 2022 Dec 16;17(12):3352-3366. doi: 
            10.1021/acschembio.1c00569. Epub 2022 Jan 7.
  PUBMED    34995053
REFERENCE   2  (bases 1 to 4811)
  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/acschembio.1c00569"; journalName: "ACS Chem Biol"; date: 
            "2022-12-16- 16"; volume: "17"; issue: "12"; pages: "3352-3366"
COMMENT     Sequence Label: pCS2_hMGFP-60A
FEATURES             Location/Qualifiers
     source          1..4811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1200..1334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1237..1256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1291..1310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(2087..2105)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2088..2105)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2140..3124)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3195..3650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3282..3301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3492..3513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3676..3780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3781..3849,3850..4641)
                     /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     complement(3999..4018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaac
     1081 gtctcttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt
     1141 ttttcctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1321 atcatgtctg gatctacgta atacgactca ctatagttct agaggctcga gttagccggc
     1381 ctggcggggt agtccgctgt gggcttcggc gtgctcgtac agcttgactt tgttgtagtc
     1441 cttgtcgtgg ctcacgatct cgatgcggtg gtccacgaag tggtagtcgg gaagctgcac
     1501 caccttcttg gctttgtagg tggtcttgaa gtcgcagcgg tagtggccgc ctccctccaa
     1561 gagcagtgcc atattaacat cgcccttcag tacgccgtcc cgcacgtaca tcttctcggt
     1621 actgggctcc cactttaggg tcttgcgctg catcaccggg ccattagcag ggaagttgac
     1681 cccgtcgaag cggattttgt acacgaagca gtcgtccaca cccttcatca tggtgatgtc
     1741 gtttgtagcg atacagattc cctggtcctc gtatgtcatg cttcgctccc acgagtagcc
     1801 ctcggggaag gtctgcttga agtagtcagg gatgtccttg gggtacttgg cgaagacacg
     1861 gttaccgtag tcgaacacgg tggtgagaat gtcataagcg aagggcaggg gggcgccctc
     1921 gatcacggtc aggtccatag tctgcttacc ctcaaagggc ttgcctttcc cgtcgccctc
     1981 gatcacgaat ttgtggccgt tcacggcgcc ctccatccgc agcttgatct tcatgtcggg
     2041 cttgatcacg cccatggatc ctgcaaaaag aacaagtagc ttgtattcta tagtgtcacc
     2101 taaatcaagc ttgctccgac gtccccaggc agaatggcgg ttccctaaac gagcattgct
     2161 tatatagacc tcccattagg cacgcctacc gcccatttac gtcaatggaa cgcccatttg
     2221 cgtcattgcc cctccccatt gacgtcaatg gggatgtact tggcagccat cgcgggccat
     2281 ttaccgccat tgacgtcaat gggagtactg ccaatgtacc ctggcgtact tccaatagta
     2341 atgtacttgc caagttacta ttaatagata ttgatgtact gccaagtggg ccatttaccg
     2401 tcattgacgt caataggggg cgtgagaacg gatatgaatg ggcaatgagc catcccattg
     2461 acgtcaatgg tgggtggtcc tattgacgtc aatgggcatt gagccaggcg ggccatttac
     2521 cgtaattgac gtcaatgggg gaggcgccat atacgtcaat aggaccgccc atatgacgtc
     2581 aataggaaag accatatata gagaccattg acgtcaatgg gggagtggct atgggcggta
     2641 ttaggaagcc ccatatatgg tatatgggac cgcccattgg gaggggctat ctacgtcaat
     2701 aggaaaaccc atatatggaa tactatatgg catagggcca atacatagta ttgaacctgg
     2761 ccaatagcca tattggcata gggccatatt ggatattgcc tatatattga tcctggcata
     2821 tagccaatat ggccgccatt attggcacca tgccaattca atatggcgga cttggcacag
     2881 ggccaagacc cctcctcaaa cttggcaccg tgccaagtag acccctcccc agttggcaca
     2941 gtgccaggtc cgccatattg aattggcatg gtgccaagtc cgccatattg aattggctgg
     3001 ggtccagatc caccatattg aattggcacg agtccatata cgccatattg aattggcaca
     3061 ggtccagatc caccatattg aattggcatg agtccatata cgccatattg aattggcggt
     3121 cgactggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3181 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3241 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3301 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3361 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3421 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3481 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3541 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3601 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     3661 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3721 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3781 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3841 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3901 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3961 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4021 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4081 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4141 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4201 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4261 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4321 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4381 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4441 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4501 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4561 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4621 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4681 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4741 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4801 aaaggatctt c
//
