LOCUS       sequence-251848-        3359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fuGFP constitutively in E.coli cells. Designed to act as 
            modular template for cutting out or amplifying fuGFP to place in 
            other plasmid backbones.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3359)
  TITLE     A new open-source superfolding green fluorescent protein: Free-Use 
            GFP (fuGFP)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3359)
  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            "Unpublished"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: pUS252
FEATURES             Location/Qualifiers
     source          1..3359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..46
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    163..184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(199..204,205..222,223..229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    237..253
                     /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     complement(1047..1064)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1047..1063)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1056..1078)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1568..2362
                     /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(1622..1641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2232..2251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2646..3234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3135..3154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
       61 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      121 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      181 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      241 gagcggataa caatttcacg gatccgaatt ctagacctcg aggaggtttc atatggtgag
      301 cagtggcgaa gatattttct cgggcttggt tccgattctg atcgagctgg agggcgatgt
      361 gaacggtcat cgttttagcg ttcgcggtga aggttatggc gacgcgagca acggcaaact
      421 ggaaattaag ttcatctgca cgaccggtcg cctgccggtg ccttggccga ccttggtgac
      481 gaccttgtcg tatggcgtgc agtgttttgc gaagtatccg gagcacatgc gccaaaacga
      541 tttctttaaa agtgcgatgc cggacggtta cgtccaggag cgtaccattt ccttcaagga
      601 agatggcacg tacaaaactc gcgcagaggt taagtttgaa ggtgaagcgc tggtcaatcg
      661 tatcgatttg aagggtttgg agtttaaaga ggatggtaac attctgggcc ataaactgga
      721 gtatagcttc aacagccatt atgtttacat tacggcagac aagaatcgta acggcttgga
      781 ggcccaattc cgtattcgcc acaatgttga tgacggtagc gtccaactgg ccgaccatta
      841 ccaacagaac accccaattg gtgagggtcc ggtgttgctg ccggaacaac actatctgac
      901 gaccaatagc gtgctgagca aagatccgca ggaacgccgt gatcacatgg tcctggtgga
      961 atttgtgacc gctgcgggct tgagcctggg tatggacgag ctgtataaga gctaatgaaa
     1021 gcttgctgca ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     1081 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     1141 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     1201 tggcgataag ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa
     1261 gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     1321 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     1381 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     1441 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     1501 gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc
     1561 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     1621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     1681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     1741 tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     1801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     1861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     1921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     1981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     2041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat
     2101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2341 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2401 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2461 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2521 ttcttcgccc accccaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2761 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3301 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatac
//
