LOCUS       sequence-269406-        2861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mNeonGreen monomeric fluorescent protein expression for mammalian 
            cells under EF1a-HTLV promoter in minimal backbone plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2861)
  TITLE     Constitutive promoter minimal backbone plasmids for optimized 
            expression in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2861)
  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            "Unpublished"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: EF1-alpha-mNeonGreen-Z1
FEATURES             Location/Qualifiers
     source          1..2861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..272
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             285..553
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      617..626
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             623..1333
                     /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      878..887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1385..1506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1396..1415)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1450..1469
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1665..2039)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(2049..2067)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2059..2105)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(2178..2766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2258..2277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccgcctg cagaagcttt actaactagt ggatctgcga tcgctccggt gcccgtcagt
       61 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
      121 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      181 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      241 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
      301 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
      361 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
      421 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
      481 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
      541 gccgttacag atccaagctg tgaccggcgc ctacaattct gcagtcgacg gtaccgcggg
      601 cccgggatcc accggtgcca ccatggtgag caagggcgag gaggataaca tggcctctct
      661 cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt
      721 gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa
      781 gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca
      841 gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg
      901 ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg
      961 ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc
     1021 tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa
     1081 gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa
     1141 tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc
     1201 taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa
     1261 gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga
     1321 gctgtacaag taataggcgg ccgcgactct agatgtacac atatggctag ctggccagac
     1381 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1441 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1501 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1561 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg aattctggac tcacaaagaa
     1621 aaaacgcccg gtgtgcaaga ccgagcgttc tgaacaattt atcctcagtc ctgctcctct
     1681 gccacaaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc
     1741 tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc
     1801 tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg
     1861 tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca
     1921 ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg
     1981 accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg
     2041 atggccctcc tatagtgagt cgtattatac tatgccgata tactatgccg atgattaatt
     2101 gtcaaggatc ctaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2161 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2641 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2761 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtgaaa actcacgtta
     2821 agggattttg gtcatggcta gttaattaac atttaaatca g
//
