LOCUS       sequence-415532-        4871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a protein of interest fused to the C-terminus of StayGold
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4871)
  TITLE     To express a protein of interest fused to the C-terminus of StayGold
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4871)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCS2+StayGold-C Cloning Vector
FEATURES             Location/Qualifiers
     source          1..4871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..985
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1020..1038
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1020..1037
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1117..1794
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSKGEELFTGVTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTL
                     LYPLLSDKSKCVEAYQNTIIKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     polyA_signal    1908..2042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1932..1951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1986..2005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2155..2175)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2155..2173)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2194..2210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2207..2229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2218..2234
                     /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(2242..2248,2249..2266,2267..2272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2287..2308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2425..2442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2596..3184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2676..2695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3355..4146,4147..4215))
                     /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     3978..3997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4216..4320)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4346..4801)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4483..4504)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4695..4714
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
       61 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
      121 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
      181 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
      241 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
      301 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
      361 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
      421 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
      481 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
      541 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
      601 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
      661 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
      721 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
      781 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
      841 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
      901 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
      961 tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga
     1021 tttaggtgac actatagact tgttcttttt gcaggatcta ccatgctgct gcccgtcccc
     1081 ctgctgctgg gcctgctggg cgccgccgcg gatccgatgg tgagcaaggg cgaggagctg
     1141 tttacaggcg tgaccccctt caagttccag ctgaagggca ccatcaacgg caagagcttc
     1201 accgtggaag gcgagggcga gggcaatagc cacgagggca gccacaaagg caagtacgtg
     1261 tgcaccagcg gcaaactgcc aatgtcttgg gccgccctgg gaactagctt cggctatggc
     1321 atgaagtact acaccaagta ccccagcggc ctgaagaact ggttccacga ggtgatgccc
     1381 gagggcttca cctacgacag acacatccag tacaagggcg acggcagcat ccacgccaag
     1441 caccagcact tcatgaagaa cggcacctac cacaacatcg tggagttcac cggccaggac
     1501 ttcaaggaga acagccccgt gctgaccggc gacatgaacg tgagcctgcc caacgaggtg
     1561 cagcacatcc ccagagatga cggcgtggag tgcccagtga ccctgctgta ccctctgctg
     1621 agcgacaaga gcaagtgcgt ggaggcctac cagaacacca tcatcaagcc cctgcacaat
     1681 cagccagccc ccgatgtgcc ataccactgg atcagaaagc agtacaccca gagcaaggac
     1741 gacaccgagg agagagacca catcatccag agcgagaccc tggaggccca cctgccatgg
     1801 cacgagcctt ctgcttctgc cgtgaaggat gaactgggat ctgagctcca tcgattcgaa
     1861 ttcaaggcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1921 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1981 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2041 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2101 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     2161 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2221 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     2281 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2341 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2401 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3001 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4321 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4381 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4441 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4501 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4561 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4621 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4681 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4741 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4801 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4861 ctattacgcc a
//
