LOCUS       Exported                3313 bp ds-DNA     circular SYN 18-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
FEATURES             Location/Qualifiers
     source          1..3313
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          283..290
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          323..444
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1067..1074
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    187..282
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    323..444
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    463..554
                     /label=RNAP II pause site
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     promoter        574..946
                     /label=lexA Operator
                     /note="LexA Operator"
     misc_feature    637..671
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     misc_feature    673..707
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     misc_feature    714..748
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     misc_feature    750..784
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     misc_feature    792..826
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     misc_feature    828..862
                     /label=ColE1 Operator
                     /note="ColE1 Operator"
                     /note="ColE1 operator sequences (Ebina et al., 1983)"
     promoter        959..1051
                     /label=cFos min pro
                     /label=cFos
                     /note="mmu-cFos min pro"
                     /note="murine c-Fos minimal promoter"
     protein_bind    1081..1205
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1223..1239)
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1247..1263
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(1271..1301)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1316..1337
                     /label=E. coli catabolite activator protein binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1625..2213)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2391..3200)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=aph(3')-Ia
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
      241 agcaatgctt ttttataatg ccaactttgt atagaaaagt tggcggccgc tacgtagttt
      301 aaacggtacc ccatggctcg agaacttgtt tattgcagct tataatggtt acaaataaag
      361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
      421 gtccaaactc atcaatgtat cttagatatc gctagcagta ctaacatacg ctctccatca
      481 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
      541 cagaacattt ctctcagatc tatcgatacg cgtttgcatg ccgcttgggc tgcaggtcga
      601 ggctaaaaaa ctaatcgcat tatcatcccc tcgacgtact gtacatataa ccactggttt
      661 tatatacagc agtactgtac atataaccac tggttttata tacagcatct agatactgta
      721 catataacca ctggttttat atacagcagt actgtacata taaccactgg ttttatatac
      781 agcagaattc gtactgtaca tataaccact ggttttatat acagcagtac tgtacatata
      841 accactggtt ttatatacag cagtcgaggt aagattagat atggatatgt atatggatat
      901 gtatatggtg gtaatgccat gtaatatgct cgactctagg atcttcgtcg acggatcccc
      961 agtgacgtag gaagtccatc cattcacagc gcttctataa aggcgccagc tgaggcgcct
     1021 actactccaa ccgcgactgc agcgagcaac taacactagt aagcttttaa ttaatacgta
     1081 tcaagtttgt acaaaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
     1141 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca
     1201 ctatgagctt ggtgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1261 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1321 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1381 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1441 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1561 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2041 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2281 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2341 ttaaatcaag cccaatctga ataatgttac aaccaattaa ccaattctga ttagaaaaac
     2401 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     2461 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2521 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2581 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2641 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2701 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2761 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2821 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2881 acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2941 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3001 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3061 gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga
     3121 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt
     3181 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     3241 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
     3301 ggccagagct gca
//
