LOCUS       sequence-57365-e        5919 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 5919)
  AUTHORS   Jang CW, Magnuson T
  TITLE     A Novel Selection Marker for Efficient DNA Cloning and 
            Recombineering in E. coli.
  JOURNAL   PLoS One. 2013;8(2):e57075. doi: 10.1371/journal.pone.0057075. Epub 
            2013 Feb 20.
  PUBMED    23437314
REFERENCE   2  (bases 1 to 5919)
  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 One.";
            date: "2013"; volume: "8(2):e57075. doi"; pages: " 
            10.1371/journal.pone.0057075. Epub 2013 Feb 20"
COMMENT     Sequence Label: pF2-DTA
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     787..809
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     801..818
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     802..818
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     825..844
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        825..843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    926..959
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1088..2266
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1319..2257
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2214..2234
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     2475..2494
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2569..3141
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          3218..3283
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     primer_bind     complement(3351..3368)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3357..3581
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3628..3646)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3667..3683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3667..3683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3680..3702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3691..3707
                     /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)."
     promoter        complement(join(3715..3721,3722..3739,3740..3745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3760..3781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3898..3915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4069..4657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4149..4168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccctttca tggcgatggc tgggacggtt cagaccttgc cgaatattct ccagcaccgt
       61 ctccatgttt tcaccacaac agctattcgg ctcggtctca gggttattgt ctcatgagcg
      121 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      181 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      241 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      301 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      361 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      421 gttcacgtag tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      481 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      541 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      601 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      661 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      721 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      781 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
      841 agggcgaatt gggtaccggg ccccccctcg aggccggcct taagacgtcg cgagatctca
      901 attgtacatc gattcccggg ctagcataac ttcgtataat gtatgctata cgaagttatc
      961 cggatgcatc tagatatccg cggtcgacaa gcttgaattc gcggccgctt aattaaggcg
     1021 cgccgtttag cttgatttat aaaggtagca gatctggacc ttctaggtct tgaaaggagt
     1081 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     1141 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
     1201 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     1261 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     1321 aagtgccgtg tgtggttccc gctggcctgg cctctttacg ggttatggcc cttgcgtgcc
     1381 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     1441 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     1501 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     1561 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     1621 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     1681 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     1741 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     1801 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     1861 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     1921 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     1981 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     2041 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     2101 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     2161 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     2221 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa ttagcttggt
     2281 aggatccact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     2341 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     2401 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     2461 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     2521 ttcctcatct ccgggccttt cgacctgcag gtcctcgcca tggatcctga tgatgttgtt
     2581 gattcttcta aatcttttgt gatggaaaac ttttcttcgt accacgggac taaacctggt
     2641 tatgtagatt ccattcaaaa aggtatacaa aagccaaaat ctggtacaca aggaaattat
     2701 gacgatgatt ggaaagggtt ttatagtacc gacaataaat acgacgctgc gggatactct
     2761 gtagataatg aaaacccgct ctctggaaaa gctggaggcg tggtcaaagt gacgtatcca
     2821 ggactgacga aggttctcgc actaaaagtg gataatgccg aaactattaa gaaagagtta
     2881 ggtttaagtc tcactgaacc gttgatggag caagtcggaa cggaagagtt tatcaaaagg
     2941 ttcggtgatg gtgcttcgcg tgtagtgctc agccttccct tcgctgaggg gagttctagc
     3001 gttgaatata ttaataactg ggaacaggcg aaagcgttaa gcgtagaact tgagattaat
     3061 tttgaaaccc gtggaaaacg tggccaagat gcgatgtatg agtatatggc tcaagcctgt
     3121 gcaggaaatc gtgtcaggcg atctctttgt gaaggaacct tacttctgtg gtgtgacata
     3181 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     3241 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     3301 tgatgaatgg gagcagtggt ggaatgcaga tcctagagct cgctgatcag cctcgactgt
     3361 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3421 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3481 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3541 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3601 cagctggggc tcgaccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta
     3661 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3721 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3781 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3841 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3901 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3961 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4501 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4741 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4801 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg ccgcccatct
     4861 ttaccaacag aacgattatt tcagttcgag ttcgttcatt gcagcaatgc tgaaaccgcc
     4921 gtcaacgtgg accacttcac cggagatacc ggcagagaga tcggagcaca ggaatgccgc
     4981 agagttaccc acatcttcaa tagtaacggt acggcgaatc ggggtaacgg cttcgcaatg
     5041 agccagcatt ttgcggaagt ctttgatacc ggaggccgcc agagtacgga tcggaccagc
     5101 agagatggcg ttaacacgca caccttccgg acccatcgcg ttcgccatat agcgcacgtt
     5161 cgcttccaga gacgcttttg ccagacccat aacgttgtag ttcgggatag cgcgctcagc
     5221 gccaaggtag gaaagggtca gcagggcaga acccggattc agcatggagc ggcaagcttt
     5281 tgccattgca acgaagctgt aggagctgat gtcgtgggca attttgaagc cttcacgggt
     5341 aacggcgtta acatagtcac catccagctg atcgccaggt gcaaaaacaa tagagtgtac
     5401 gaaaccgtca aatttcggcc aaactttccc cagttcagcg aacatggtgt cgatgctggc
     5461 atcttctgca acatcgcact gcagaacgat gtcagaaccc aattgagcgg caaattcttc
     5521 tacgcggcct ttcagtttgt cgttctggta ggtgaatgcc agttcagctc cttcgcggtg
     5581 catcgcctga gcgataccgt aggcgatgga tagtttgctg gcaacaccgg ttaccagaat
     5641 gcgcttaccg gaaagaaaac ccatagcttt aatccttatt gttgatgctt gttgtgcctg
     5701 aaaatcaggc gattcgtcgt tttagtaaac agtacgaaca gataaacggt tattataatc
     5761 aacctggctg tgagtagcta tagttgccag gtccgaccgg agcaggctgc ggcagggggg
     5821 gcgcttttcc cctcacccta accctctccc cagaggggcg aggggaccgt attgtgcaaa
     5881 tattgttacc ccagcaacaa acaggctcat acagcccct
//
