LOCUS       sequence-213796-        3742 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  shuttle vector, C-terminal GFP tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3742)
  AUTHORS   Paschke P, Knecht DA, Silale A, Traynor D, Williams TD, Thomason PA,
            Insall RH, Chubb JR, Kay RR, Veltman DM
  TITLE     Rapid and efficient genetic engineering of both wild type and axenic
            strains of Dictyostelium discoideum.
  JOURNAL   PLoS One. 2018 May 30;13(5):e0196809. doi: 
            10.1371/journal.pone.0196809. eCollection 2018.
  PUBMED    29847546
REFERENCE   2  (bases 1 to 3742)
  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: "2018-05-30"; volume: "13(5):e0196809. doi"; pages: " 
            10.1371/journal.pone.0196809. eCollection 2018"
COMMENT     Sequence Label: pDM1021
FEATURES             Location/Qualifiers
     source          1..3742
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1108..1125)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1108..1124)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1117..1139)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1333..1352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1452..1474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1512..1530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1598..1702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1703..1771,1772..2563)
                     /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(1921..1940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2734..3322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3223..3242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3476..3493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3610..3631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3646..3651,3652..3669,3670..3676)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3684..3700
                     /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     3689..3711
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3708..3724
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3708..3724
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tttttttttt tttttttctt atttcttaaa acaaataaat taaattaaat aaaaaataaa
       61 aatcagatct tcaactagtg gtggttcagg aggtagttca aaaggtgaag aattatttac
      121 aggtgttgtt ccaattttag ttgaattaga tggtgatgtt aatggtcata aattttcagt
      181 ttcaggtgaa ggtgaaggtg atgcaacata tggtaaatta acattaaaat ttatttgtac
      241 aacaggtaaa ttaccagttc catggccaac attagttaca acatttacat atggtgttca
      301 atgtttttca agatatccag atcatatgaa acaacatgat ttttttaaat cagcaatgcc
      361 agaaggttat gttcaagaaa gaacaatttt ttttaaagat gatggtaatt ataaaacaag
      421 agcagaagtt aaatttgaag gtgatacatt agttaataga attgaattaa aaggtattga
      481 ttttaaagaa gatggtaata ttttaggtca taaattagaa tataattata attcacataa
      541 tgtttatatt atggcagata aacaaaaaaa tggtattaaa gttaatttta aaattagaca
      601 taatattgaa gatggttcag ttcaattagc agatcattat caacaaaata caccaattgg
      661 tgatggtcca gttttattac cagataatca ttatttatca acacaatcag cattatcaaa
      721 agatccaaat gaaaaaagag atcatatggt tttattagaa tttgttacag cagcaggtat
      781 tacacatggt atggatgaat tatataaata agctagttaa ataaataaat tatttaataa
      841 ataataaaaa aacaaattgt tgtaataatc taatattttc tttttttttt aatttttttt
      901 ttttaaatct taataattat taagttattt taattttttt tttttttttt tttttttttt
      961 tttttttttc tatcaaaaaa atcaaatata tttaaaaaat ttattattta cagatacatt
     1021 ttgaatggtg aagataaata tatgcattag atgtaaaaca gccaaagagt atgaaaatca
     1081 aaaagatagc cggccatcaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     1141 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1201 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1261 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     1321 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     1381 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     1441 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     1501 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     1561 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1921 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2221 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2341 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2401 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2461 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2521 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2581 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2641 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3361 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3421 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3481 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3541 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3601 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     3661 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3721 tgacatgatt acgaattcga tg
//
