LOCUS       sequence-365358-        4203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amp-resistant, low copy (p15A ori), E. coli 5'ptsI transcriptionally
            fused with GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4203)
  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 4203)
  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: pDY380
FEATURES             Location/Qualifiers
     source          1..4203
                     /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             2348..3064
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2378..2406)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3001..3021
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        3144..3248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3249..3317,3318..4109)
                     /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(3467..3486)
                     /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 gtcataatcc acgagatgcg gcccaattta ctgcttagga
     2341 gaagatcatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     2401 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     2461 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     2521 aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat
     2581 gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
     2641 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
     2701 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     2761 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     2821 gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
     2881 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     2941 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     3001 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     3061 atagaatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3181 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3241 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3301 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3361 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3421 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3481 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3541 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3601 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3661 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3721 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3781 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3841 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3901 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3961 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4021 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4081 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4201 atc
//
