LOCUS       sequence-248605-        4805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a protein of interest fused to the C-terminus of 
            mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4805)
  TITLE     To express a protein of interest fused to the C-terminus of 
            mNeonGreen
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4805)
  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            "Unpublished"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+mNeonGreen-C Cloning Vector
FEATURES             Location/Qualifiers
     source          1..4805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             78..785
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      333..342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    857..991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(881..900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     935..954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1104..1124)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1104..1122)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1143..1159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1143..1159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1156..1178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1167..1183
                     /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(1191..1197,1198..1215,1216..1221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1236..1257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1374..1391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1545..2133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1625..1644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2304..3095,3096..3164))
                     /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     2927..2946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3165..3269)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3295..3750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3432..3453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3644..3663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3821..4805
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg gtgagcaagg gcgaggagga taacatggcc tctctcccag
      121 cgacacatga gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc
      181 agggcaccgg caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg
      241 acctccagtt ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc
      301 tgccctaccc tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggatacc
      361 aagtccatcg cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca
      421 cctacgaggg aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg
      481 acggtcctgt gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt
      541 accccaacga caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca
      601 agcgctaccg gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact
      661 atctgaagaa ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg
      721 agctcaactt caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctgt
      781 acaagggatc tggatcccat cgattcgaat tcaaggcctc tcgagcctct agaactatag
      841 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
      901 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      961 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1021 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg
     1081 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1201 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1981 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2581 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     3301 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3361 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3421 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3481 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3541 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3601 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3661 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3721 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     3781 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat
     3841 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat
     3901 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
     3961 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag
     4021 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
     4081 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
     4141 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
     4201 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
     4261 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
     4321 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
     4381 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
     4441 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
     4501 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
     4561 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg
     4621 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
     4681 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
     4741 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
     4801 ggaac
//
