LOCUS       sequence-459279-        4503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP from jellyfish Aquorea sp. under strong synthetic constitutive 
            promoter pFAB4026; kanamycin resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  AUTHORS   Siddall A, Williams AA, Sanders J, Denton JA, Madden D, Schollar J, 
            Bryk J
  TITLE     UNIGEMS: plasmids and parts to facilitate teaching on assembly, gene
            expression control and logic in E. coli
  JOURNAL   bioRxiv 2021.06.20.449138
REFERENCE   2  (bases 1 to 4503)
  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: "bioRxiv 
            2021.06.20.449138"
COMMENT     Sequence Label: p006-GFP-strong
FEATURES             Location/Qualifiers
     source          1..4503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1083
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(20..39)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1169..1757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1658..1677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1911..1928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2026..2043
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2027..2043
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2507..2515
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2522..3238
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTRVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKVRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     3175..3195
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3434..3453)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3520..3611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3612..4421
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 atgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
       61 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
      121 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
      181 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
      241 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
      301 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
      361 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
      421 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
      481 ttctcccatg aggacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
      541 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
      601 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
      661 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      721 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      781 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agatagctca
      841 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      901 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgcca
      961 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     1021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1081 tgactcatga ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag
     1141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1321 tagtgtagcc gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg
     1381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1861 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1921 tgagcgagga agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc
     1981 ccctgacgga tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc
     2041 agtcttaagc tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata
     2101 acgtaaaaac ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt
     2161 cagaatattt aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa
     2221 atgagaaaaa agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct
     2281 ataattaaac tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt
     2341 taaagagaat taagaaaata aatctcgaaa ataataaagg gaaaatcaga aaaaatttat
     2401 ttgcttatta atcatccggc tcgtataatg tgtggagggc ccaagttcac ttaaaaagga
     2461 gatcaacaat gaaagcaatt ttcgtactga aacatcttaa tcatgctaag gaggttttct
     2521 aatgagtaaa ggagaagaac ttttcactag agttgtccca attcttgttg aattagacgg
     2581 tgatgttaat gggcacaaat tttctgtcag tggagaggga gaaggtgatg caacatacgg
     2641 aaaacttacc cttaaattta tttgcactac tggaaaacta ccagttccgt ggccaacact
     2701 tgtcactact ctctcttatg gtgttcaatg cttttcgaga tacccagatc acatgaaacg
     2761 gcatgacttt ttcaagagtg ccatgcccga gggttatgta caggaaagaa ctatattttt
     2821 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     2881 taatagaatc gagttaaaag gtgttgattt taaagaagat ggaaacattc ttggacacaa
     2941 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     3001 aatcaaagct aacttcaaag ttagacacaa cattgaagat ggaagcgttc aactggcaga
     3061 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     3121 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     3181 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataacc
     3241 ccaagggcga caccccctaa ttagcccggg cgaaaggccc agtctttcga ctgagccttt
     3301 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagtcccca cactaccatc
     3361 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
     3421 gccgccaggc aaacaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa
     3481 tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac
     3541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3601 aaaggaagaa tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     3661 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     3721 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3781 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3841 tgccacttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3901 ctgcgatccc cggaaaaaca gcgttccagg tattagaaga atatcctgat tcaggtgaaa
     3961 atattgttga tgcgctggca gtgttcctgc gccggttgca ctcgattcct gtttgtaatt
     4021 gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg
     4081 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     4141 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     4201 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     4261 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     4321 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     4381 ataaattgca atttcatttg atgctcgatg agtttttcta agcggcgcgc catcgaatgg
     4441 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     4501 aat
//
