LOCUS       sequence-382124-        3359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fuGFPs constitutively in E.coli cells. Designed to act as 
            modular template for cutting out or amplifying fuGFPs 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  JOURNAL   Unpublished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            "Unpublished"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: pUS252s
FEATURES             Location/Qualifiers
     source          1..3359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..91
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    208..229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(244..249,250..267,268..274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    282..298
                     /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(1092..1109)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1092..1108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1101..1123)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1613..2407
                     /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(1667..1686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2277..2296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2691..3279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3180..3199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
       61 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
      121 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      181 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      241 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      301 tcacggatcc gaattctaga cctcgaggag gtttcatatg gtgagcagtg gcgaagatat
      361 tttctcgggc ttggttccga ttctgatcga gctggagggc gatgtgaacg gtcatcgttt
      421 tagcgttcgc ggtgaaggtt atggcgacgc gagcaacggc aaactggaaa ttaagttcat
      481 ctgcacgacc ggtcgcctgc cggtgccttg gccgaccttg gtgacgacct tgtcgtatgg
      541 cgtgcagtgt tttgcgaagt atccggagca catgcgccaa aacgatttct ttaaaagtgc
      601 gatgccggac ggttacgtcc aggagcgtac catttccttc aaggaagatg gcacgtacaa
      661 aactcgcgca gaggttaagt ttgaaggtga agcgctggtc aatcgtatcg atttgaaggg
      721 tttggagttt aaagaggatg gtaacattct gggccataaa ctggagtata gcttcaacag
      781 ccattatgtt tacattacgg cagacaagaa tcgtaacggc ttggaggccc aattccgtat
      841 tcgccacaat gttgatgacg gtagcgtcca actggccgac cattaccaac agaacacccc
      901 aattggtgag ggtccggtgt tgctgccgga acaacactat ctgacgatca atagcgtgct
      961 gagcaaagat ccgcaggaac gccgtgatca catggtcctg gtggaatttg tgaccgctgc
     1021 gggcttgagc ctgggtatgg acgagctgta taagagctaa tgaaagcttg ctgcaggcat
     1081 gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1141 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1201 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ataagctagc
     1261 ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     1321 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     1381 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     1441 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     1501 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     1561 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1621 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1681 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1741 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tccaagacga
     1801 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1861 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1921 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1981 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2041 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2101 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg acggcgagga
     2161 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2221 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2281 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2341 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2401 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2461 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2521 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccc
//
