LOCUS       pME_Cp_0_7-8_006_Kan/ColEI_sfGFP        2808 bp    DNA     circular UNA 07-OCT-2025
DEFINITION  Kan-ColE1, with sfGFP Placeholder, only compatible with new
            overhang system, reversed backbone (same orientation as
            pMW_Cp_0_7-8_002).
ACCESSION   urn.local.burgi.39z-jqvwi8t
VERSION     urn.local.burgi.39z-jqvwi8t
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     promoter        14..143
                     /ApEinfo_fwdcolor="#9EFF91"
                     /ApEinfo_revcolor="#9EFF91"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="BBa_J72163 GlpT Promoter"
     RBS             144..179
                     /ApEinfo_fwdcolor="#9EFF91"
                     /ApEinfo_revcolor="#9EFF91"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="sfGFP Ribosome Binding Site"
     CDS             180..896
                     /ApEinfo_fwdcolor="#34FF03"
                     /ApEinfo_revcolor="#34FF03"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="sfGFP"
     terminator      897..1025
                     /ApEinfo_fwdcolor="#9EFF91"
                     /ApEinfo_revcolor="#9EFF91"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="BBa_B0015 Terminator"
     primer_bind     complement(1015..1036)
                     /created_by="primer3"
                     /Tm_(binding_region)="57.3"
                     /Tm_(with_extension)="69.5"
                     /%GC_(binding_region)="45.5"
                     /%GC_(with_extension)="45.7"
                     /Hairpin_Tm_(with_extension)="65.5"
                     /Self_Dimer_Tm_(with_extension)="41.0"
                     /annotation_group="19763272035980743d868d1-33b0-4bb7-9121-
                     268576891629"
                     /Sequence="GTCTTGAGACGTATAAACGCAG"
                     /Mismatches=0
                     /Extension="AACACCTGCATAC"
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP"
                     /label="sfGFP_rev"
     primer_bind     1037..1052
                     /created_by="primer3"
                     /Tm_(binding_region)="60.8"
                     /Tm_(with_extension)="74.5"
                     /%GC_(binding_region)="68.8"
                     /%GC_(with_extension)="57.6"
                     /Hairpin_Tm_(with_extension)="45.3"
                     /Self_Dimer_Tm_(with_extension)="37.7"
                     /annotation_group="197632592289084aabc9693-af3c-4b96-a100-
                     bcb433c9ae4f"
                     /Sequence="TGCGGCCGCGTGTTAC"
                     /Mismatches=0
                     /Extension="AACACCTGCATACAGAC"
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP"
                     /label="Kan_BB_rev"
     terminator      complement(1046..1072)
                     /ApEinfo_fwdcolor="#93FF35"
                     /ApEinfo_revcolor="#93FF35"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="KanR Terminator"
     CDS             complement(1073..1888)
                     /ApEinfo_fwdcolor="#008000"
                     /ApEinfo_revcolor="#008000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="KanR"
     primer_bind     complement(1087..1106)
                     /%GC="45.0"
                     /Hairpin_Tm="34.1"
                     /Self_Dimer_Tm="None"
                     /Tm="56.0"
                     /annotation_group="199890415603899beb5008c-a059-4972-8b49-
                     a1f75a87f902"
                     /created_by="primer3"
                     /Sequence="GCAGTTTCATTTGATGCTCG"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP PCR
                     Product 2 (reversed) - pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP
                     PCR Product"
                     /label="oMCB021_lvl2_seq_primer_fwd"
     promoter        complement(1889..2036)
                     /ApEinfo_fwdcolor="#93FF35"
                     /ApEinfo_revcolor="#93FF35"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="KanR Promoter"
     rep_origin      2113..2701
                     /note="origin of replication from pBR322 (also known as
                     pMB1, pUC origin)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="ColE1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2ht-fvq5kj1"">ColE1</a>"
                     /Alternative_Match="71.50%; Adjusted-Interval-0:3/0; <a
                     href=""urn:local:.:2ja-fvq5kj7"">p15A</a>"
                     /Alternative_Match="66.33%; Adjusted-Interval-0:2836/2743;
                     <a href=""urn:local:.:2hn-fvq5kj0"">colA</a>"
                     /label="ColE1"
     primer_bind     2718..2736
                     /%GC="52.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="56.5"
                     /annotation_group="1998904493576005b1b67ca-50b3-4d90-a39d-
                     710ce15fce42"
                     /created_by="primer3"
                     /Sequence="CCTTTTTACGGTTCCTGGC"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP PCR
                     Product 2 (reversed) - pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP
                     PCR Product"
                     /label="oMCB022_lvl2_seq_primer_rev"
     primer_bind     complement(2797..1)
                     /created_by="primer3"
                     /Tm_(binding_region)="59.6"
                     /Tm_(with_extension)="74.5"
                     /%GC_(binding_region)="84.6"
                     /%GC_(with_extension)="62.1"
                     /Hairpin_Tm_(with_extension)="52.0"
                     /Self_Dimer_Tm_(with_extension)="49.7"
                     /annotation_group="1976323988637978ada4c3f-363f-40d2-ada8-
                     2b8fc0c38696"
                     /Sequence="TGCGGCCGCACGG"
                     /Mismatches=0
                     /Extension="AACACCTGCATACTGT"
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="pMW_Cp_0_7-8_004_Kan/ColE1_v2_sfGFP"
                     /label="Kan_bb_fwd"
ORIGIN      
        1 aacatgagac gaggaaagtg aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc
       61 ttatttaata atgtgtgcgg caattcacat ttaatttatg aatgttttct taacatcgcg
      121 gcaactcaag aaacggcagg ttcggatctt agctactaga gaaagaggag aaatactaga
      181 tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg
      241 atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta
      301 aactgacgct gaagttcatc tgtactactg gtaaactgcc ggttccttgg ccgactctgg
      361 taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc
      421 atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta
      481 aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa
      541 accgcattga gctgaaaggc attgacttta aagaggacgg caatatcctg ggccataagc
      601 tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca
      661 ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc
      721 actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc
      781 tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc
      841 tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgaccag
      901 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      961 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     1021 ttatacgtct caagactgcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa
     1081 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1141 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1201 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1261 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1321 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1381 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1441 cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     1501 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     1561 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     1621 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     1681 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     1741 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     1801 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc
     1861 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     1921 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     1981 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat
     2041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2761 gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgc
//
