LOCUS       sequence-248628-        3648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses avGFP (Lambert 2019 clone) constitutively in E. coli (most
            strains).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3648)
  AUTHORS   Lambert GG, Depernet H, Gotthard G, Schultz DT, Navizet I, Lambert 
            T, Adams SR, Torreblanca-Zanca A, Chu M, Bindels DS, Levesque V, 
            Nero Moffatt J, Salih A, Royant A, Shaner NC
  TITLE     Aequorea's secrets revealed: New fluorescent proteins with unique 
            properties for bioimaging and biosensing.
  JOURNAL   PLoS Biol. 2020 Nov 2;18(11):e3000936. doi: 
            10.1371/journal.pbio.3000936. eCollection 2020 Nov.
  PUBMED    33137097
REFERENCE   2  (bases 1 to 3648)
  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: "PLoS 
            Biol."; date: "2020-11-2"; volume: "18(11):e3000936. doi"; pages: " 
            10.1371/journal.pbio.3000936. eCollection 2020 Nov"
COMMENT     Sequence Label: pNCST-avGFP
FEATURES             Location/Qualifiers
     source          1..3648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1049..1067)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1063..1110
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1207..1662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1294..1313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1504..1525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1688..1792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1793..1861,1862..2653)
                     /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     complement(2011..2030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2824..3412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3313..3332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3566..3583
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taacgatccg ctcgaggaaa acctgtactt ccagggtatg tctaagggtg aagaactgtt
      301 caccggtgtt gttcctatcc tggttgaact ggacggtgac gttaacggtc agaagttctc
      361 tgtttctggt gaaggtgaag gtgacgctac ctacggtaag ctgaccctga agttcatctg
      421 caccaccggt aagctgcctg ttccttggcc taccctggtt accaccttct cttacggtgt
      481 tcagtgcttc tctcgttacc ctgaccatat gaagcagcat gacttcttca agtctgctat
      541 gcctgaaggt tacgttcagg aacgtaccat cttctacaag gacgacggta actacaagac
      601 ccgtgctgaa gttaagttcg aaggtgacac cctggttaac cgtatcgaac tgaagggtat
      661 cgacttcaag gaagacggta acatcctggg tcataagatg gaatacaact acaactctca
      721 taacgtttac atcatggctg acaagcctaa gaacggtatc aaggttaact tcaagatccg
      781 tcataacatc aaggacggtt ctgttcagct ggctgaccat taccagcaga acacccctat
      841 cggtgacggt cctgttctgc tgcctgacaa ccattacctg tctacccagt ctgctctgtc
      901 taaggaccct aacgaaaagc gtgaccatat gatcctgctg gaattcgtta ccgctgctgg
      961 tatcacccat ggtatggacg aactgtacaa gtgacgtttg atccggctgc taacaaagcc
     1021 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1081 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc
     1141 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1201 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1261 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1321 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1381 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1441 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1501 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1561 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1621 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1681 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1741 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1861 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1921 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1981 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2041 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2101 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2161 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2221 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2281 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2341 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2401 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2461 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2521 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2581 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2641 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2701 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2761 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2821 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2881 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2941 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3001 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3061 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3121 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3181 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3241 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3301 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3361 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3421 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3481 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3541 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3601 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcag
//
