LOCUS       sequence-365359-        5085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amp-resistant, low copy (p15A ori), E. coli ptsI transcriptionally 
            fused with GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5085)
  AUTHORS   Yang DD, Rusch LM, Widney KA, Morgenthaler AB, Copley SD
  TITLE     Synonymous edits in the Escherichia coli genome have substantial and
            condition-dependent effects on fitness.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jan 30;121(5):e2316834121. doi: 
            10.1073/pnas.2316834121. Epub 2024 Jan 22.
  PUBMED    38252823
REFERENCE   2  (bases 1 to 5085)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-01-30"; volume: "121(5):e2316834121. 
            doi"; pages: " 10.1073/pnas.2316834121. Epub 2024 Jan 22"
COMMENT     Sequence Label: pDY390
FEATURES             Location/Qualifiers
     source          1..5085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      71..616
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     517..536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     733..750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3230..3946
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3260..3288)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3883..3903
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        4026..4130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4131..4199,4200..4991)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4349..4368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccttaataa
       61 gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa
      121 ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta
      181 actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc
      241 atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt
      301 gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
      361 tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt
      421 gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa
      481 ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag
      541 tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg
      601 gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc
      661 ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga
      721 acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc
      781 tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct
      841 catcagtgcc aacatagtaa gccagtatac actccgctag cgcactttcg cccctcctgg
      901 cattggttca gcctgtcgga actggtattt aaccagacta attattttga tgcgcgaaat
      961 taatcgttac aggaaaagcc aaagctgaat cgattttatg atttggttca attcttcctt
     1021 tagcggcata atgtttaatg acgtacgaaa cgtcagcggt caacacccgc cagcaatgga
     1081 ctgtattgcg ctcttcgtgc gtcgcgtctg ttaaaaactg gcgctaacaa tacaggctaa
     1141 agtcgaaccg ccaggctaga ctttagttcc acaacactaa acctataagt tggggaaata
     1201 caatgttcca gcaagaagtt accattaccg ctccgaacgg tctgcacacc cgccctgctg
     1261 cccagtttgt aaaagaagct aagggcttca cttctgaaat tactgtgact tccaacggca
     1321 aaagcgccag cgcgaaaagc ctgtttaaac tgcagactct gggcctgact caaggtaccg
     1381 ttgtgactat ctccgcagaa ggcgaagacg agcagaaagc ggttgaacat ctggttaaac
     1441 tgatggcgga actcgagtaa tttcccgggt tcttttaaaa atcagtcaca agtaaggtag
     1501 ggttatgatt tcaggcattt tagcatcccc gggtatcgct ttcggtaaag ctctgcttct
     1561 gaaagaagac gaaattgtca ttgaccggaa aaaaatttct gccgaccagg ttgatcagga
     1621 agttgaacgt tttctgagcg gtcgtgccaa ggcatcagcc cagctggaaa cgatcaaaac
     1681 gaaagctggt gaaacgttcg gtgaagaaaa agaagccatc tttgaagggc atattatgct
     1741 gctcgaagat gaggagctgg agcaggaaat catagccctg attaaagata agcacatgac
     1801 agctgacgca gctgctcatg aagttatcga aggtcaggct tctgccctgg aagagctgga
     1861 tgatgaatac ctgaaagaac gtgcggctga cgtacgtgat atcggtaagc gcctgctgcg
     1921 caacatcctg ggcctgaaga ttatcgacct gagcgccatt caggatgaag tcattctggt
     1981 tgccgctgac ctgacgccgt ccgaaaccgc acagctgaac ctgaagaagg tgctgggttt
     2041 catcaccgac gcgggtggcc gtacttccca cacctctatc atggcgcgtt ctctggaact
     2101 acctgctatc gtgggtaccg gtagcgtcac ctctcaggtg aaaaatgacg actatctgat
     2161 tctggatgcc gtaaataatc aggtttacgt caatccaacc aacgaagtta ttgataaaat
     2221 gcgcgctgtt caggagcaag tggcttctga aaaagcagag cttgctaaac tgaaagatct
     2281 gccagctatt acgctggacg gtcaccaggt agaagtatgc gctaacattg gtacggttcg
     2341 tgacgttgaa ggtgcagagc gtaacggcgc tgaaggcgtt ggtctgtatc gtactgagtt
     2401 cctgttcatg gaccgcgacg cactgcccac tgaagaagaa cagtttgctg cttacaaagc
     2461 agtggctgaa gcgtgtggct cgcaagcggt tatcgttcgt accatggaca tcggcggcga
     2521 caaagagctg ccatacatga acttcccgaa agaagagaac ccgttcctcg gctggcgcgc
     2581 tatccgtatc gcgatggatc gtagagagat cctgcgcgat cagctccgcg ctatcctgcg
     2641 tgcctcggct ttcggtaaat tgcgcattat gttcccgatg atcatctctg ttgaagaagt
     2701 gcgtgcactg cgcaaagaga tcgaaatcta caaacaggaa ctgcgcgacg aaggtaaagc
     2761 gtttgacgag tcaattgaaa tcggcgtaat ggtggaaaca ccggctgccg caacaattgc
     2821 acgtcattta gccaaagaag ttgatttctt tagtatcggc accaatgatt taacgcagta
     2881 cactctggca gttgaccgtg gtaatgatat gatttcacac ctttaccagc caatgtcacc
     2941 gtccgtgctg aacttgatca agcaagttat tgatgcttct catgctgaag gcaaatggac
     3001 tggcatgtgt ggtgagcttg ctggcgatga acgtgctaca cttctgttgc tggggatggg
     3061 tctggacgaa ttctctatga gcgccatttc tatcccgcgc attaagaaga ttatccgtaa
     3121 cacgaacttc gaagatgcga aggtgttagc agagcaggct cttgctcaac cgacaacgga
     3181 cgagttaatg acgctggtta acaagttcat tgaagaaaaa acaatctgca tgagtaaagg
     3241 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     3301 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     3361 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt
     3421 cggttatggt gttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt
     3481 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
     3541 gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
     3601 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
     3661 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa
     3721 cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca
     3781 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca
     3841 atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt
     3901 aacagctgct gggattacac atggcatgga tgaactatac aaatagaata cgcctatttt
     3961 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4021 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     4381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4561 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4621 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     4681 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4741 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4801 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4861 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4921 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4981 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5041 atttttaatt taaaaggatc taggtgaaga tcctttttga taatc
//
