LOCUS       sequence-10128-e        3453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3453)
  AUTHORS   Gully D, Bouveret E
  TITLE     A protein network for phospholipid synthesis uncovered by a variant 
            of the tandem affinity purification method in Escherichia coli.
  JOURNAL   Proteomics. 2006 Jan . 6(1):282-93.
  PUBMED    16294310
REFERENCE   2  (bases 1 to 3453)
  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: "Proteomics.
            2006 Jan . 6(1):282-93."
COMMENT     Sequence Label: pEB354
FEATURES             Location/Qualifiers
     source          1..3453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(717..733)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(718..734)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(748..770)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1135..1929)
                     /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(1246..1265)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1856..1875
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      2325..2869
                     /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     2770..2789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2986..3003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3344..3365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3380..3385,3386..3403,3404..3410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3418..3434
                     /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     3423..3445
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3442..3453,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3442..3453,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatgc agcaatcgca tcaggctggt tacgcaaacg ccgccgaccg ggagtctggc
       61 atccccgcag ccgtactcga tggcatcaag gccgtggcga aggaaaaaaa cgccacattg
      121 atgttccgcc tggtcaaccc ccattccacc agcctgattg ccgaaggggt ggccaccaaa
      181 ggattgggcg tgcacgccaa gtcgtccgat tgggggttgc aggcgggcta cattcccgtc
      241 aacccgaatc tttccaaact gttcggccgt gcgcccgagg tgatcgcgcg ggccgacaac
      301 gacgtcaaca gcagcctggc gcatggccat accgcggtcg acctgacgct gtcgaaagag
      361 cggcttgact atctgcggca agcgggcctg gtcaccggca tggccgatgg cgtggtcgcg
      421 agcaaccacg caggctacga gcagttcgag tttcgcgtga aggaaacctc ggacgggcgc
      481 tatgccgtgc agtatcgccg caagggcggc gacgatttcg aggcggtcaa ggtgatcggc
      541 aatgccgccg gtattccact gacggcggat atcgacatgt tcgccattat gccgcatctg
      601 tccaacttcc gcgactcggc gcgcagttcg gtgaccagcg gcgattcggt gaccgattac
      661 ctggcgcgca cgcggcgggc tgcagggtcg actctagaat tcgatatcaa gcttatcgat
      721 accgtcgacc tcgagcaatt cggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      781 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      841 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctg
      901 atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca
      961 gggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     1021 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     1081 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     1141 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     1201 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     1261 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     1321 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     1381 atcctcgccg tcgggcatcc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
     1441 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     1501 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     1561 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     1621 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     1681 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     1741 ctcgtcttgg agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     1801 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     1861 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     1921 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca
     1981 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     2041 agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag
     2101 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct
     2161 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     2221 tttctacgtg ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc ggcagcgtga
     2281 agctgtgcct agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt
     2341 ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag
     2401 gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca
     2461 aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa
     2521 tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag
     2581 acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc
     2641 cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg
     2701 gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg
     2761 agccgccagg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     2821 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     2881 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     2941 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     3001 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     3061 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     3121 tatacactcc gctaagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca
     3181 ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga
     3241 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg atctggcacg
     3301 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     3361 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     3421 tgagcggata acaatttcac acaggaaaca gct
//
