LOCUS       pBTK1057_(GFP_and_KanR) 5708 bp ds-DNA     circular     20-SEP-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     primer_bind     447..466
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="note: pBR322 origin, forward primer"
     misc_feature    664..1720
                     /label="Homology Arm 1"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1748..1806
                     /label="CP25"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1817..1824
                     /label="RBS"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1833..2549
                     /label="GFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer_bind     complement(2652..2671)
                     /label="pENTR-R"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: pENTR vectors, reverse primer"
     CDS             2663..3478
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="gene: aph(3')-Ia product: aminoglycoside phosphotransferase note: confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes translation: MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2736..2755)
                     /label="Kan-R"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="note: Kanamycin resistance gene, reverse primer"
     misc_feature    3513..4486
                     /label="Homology Arm 2"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             complement(4599..5459)
                     /label="AmpR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="gene: bla product: beta-lactamase note: confers resistance to ampicillin, carbenicillin, andrelated antibiotics translation: MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     5222..5241
                     /label="Amp-R"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="note: Ampicillin resistance gene, reverse primer"
     promoter        complement(5460..5564)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="gene: bla"
     rep_origin      5667..546
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: RIGHT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccagggga aacgcctggt atctttatag tcctgtcggg
      481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc cgcccctGAA
      661 TTCaacctgc cctataccgt atggttttta ttcatggcag gctgtctgtc cctgtttctg
      721 tgtatttctt atatctatta ccgcaacaaa ttcataacaa cctacgcata cctaccctac
      781 ggacgtgcca gcaacgccga ccagcttccc aacgtcaact ggataaaatt aaccagcccc
      841 tgcctgcctg agacctctgt cattaatggc attgttgccg acctgcacga acctgagctc
      901 gacaaccact ggcagaaatt tcttgcccgg caaaccctca atggcatgcc tgtctacaat
      961 attcgtcagg tagaagaatt tcttaccgga cgggttaaaa ttcatcacct atatgaaaat
     1021 gatcttggtt ctttgctgcc ttcaccaacc tatgtattga ttaaacgcgt attcgacatc
     1081 gttcttatcc tgctatccgc ccctgttacc ctgccggtaa tgcttattac tgccattgcc
     1141 atcaaacttg aaagcccggg taacatcctg ttccggcaaa accgtgtagg acgcagcggc
     1201 aaagaattct gtatttataa atttcgcagc atgtgcagcg attcagagaa aaacggtgcc
     1261 cgcatggcgc aacttgggga tgcccgcgtc acccgtatcg gacacatcat ccgcaaaacc
     1321 cgcatcgacg agctgccgca gttctggaac atcctgaaag gtgacatgag cctgattggt
     1381 ccccgccctg agcaaaaagt atttgtcgac cagtttattc aggaaattcc cttttatgac
     1441 taccgccata ttgtcaaacc cggtatttcc ggctgggcgc aagttactca gggctacgct
     1501 gccgacaccg aacaaaccca gataaaaatc gaacatgatt tttactacat caaacatttt
     1561 tccttctcac tcgacatact catattcttc aaaaccctca aaaccatgct caccggcttc
     1621 ggcgcacgct aacccagcca tccccataca aaacgctacc agtaaaatct taccggtagc
     1681 gttgcaatac acttacaaaa accacatagc acaactgccg AACGTGGCAA TTCGTCGCAA
     1741 TACAAcgctt tggcagttta ttcttgacat gtagtgaggg ggctggtata atcacatagt
     1801 actgttatac agaaacagag gagatattac atatgagtaa aggagaagag cttttcacag
     1861 gagttgtccc aatcctcgtg gaattagacg gtgatgttaa tgggcacaag ttctctgtca
     1921 gtggagaggg tgaaggcgac gcaacatatg gcaagctgac ccttaaattt atttgcacca
     1981 cgggtaaact acctgttcca tggccaacac tggtcactac gttcgggtat ggggttcagt
     2041 gctttgcgcg ctacccagat cacatgaaac agcacgactt tttcaagagt gcaatgcccg
     2101 aaggctatgt acaggagaga accatctttt ttaaggatga cggcaactat aagacacgcg
     2161 ccgaagtgaa gttcgagggt gatacccttg ttaatagaat cgagttaaag ggtattgact
     2221 ttaaggaaga tggaaatatt ttaggccaca aactggaata taactataac tcccataatg
     2281 tgtacattat ggccgacaag caaaagaacg gtatcaaggt taacttcaag atcagacaca
     2341 acattgagga tggaagcgtt caactagccg accattacca acaaaacacc ccaattggcg
     2401 atgggcctgt gctgttacca gacaaccatt acctgtccac tcaatctgcc ctttcgaaag
     2461 atcccaacga aaagcgcgac cacatggtcc ttcttgagtt tgtcacggct gctgggatta
     2521 cacacggcat ggatgaacta tacaaataaA TCCGTAATCG TTAATCCGCA AATCCCtctg
     2581 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2641 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc
     2701 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2761 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     2821 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2881 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2941 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     3001 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3061 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca
     3121 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3181 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     3241 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3301 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3361 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3421 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3481 agaattggtt aattggttgt aacacgcgGC TGgctgcacc agtaacgctg attatctggc
     3541 tctagaaaaa actcaaatac caaaatttat cagacacgga taaaaatgat taaacgtatt
     3601 gtaaatataa ataccttatc gaataaatgc agatgactaa tgttctgaaa aaagataagc
     3661 tacaatgaac tatcacaaac cgatgcaatg ctcagggagc cggcatgacc ctgtttctac
     3721 ccttgattta cctgctctgt ggtctgatcg ccggcaaatt aacccgggat acaaaacctg
     3781 ttttgtctct gctactgaca aaatttatta tccctgtcgt aattatctat aatatagcga
     3841 ccaatttcag caatatgggc gcggtattaa ttaccactat tgttgttatg agcataatgc
     3901 tgcttgccgg taaatttctc cggatcaatc cggtaagcac cctgtgtttc agctatctga
     3961 atattggctg gctcggtctt cccatagcag cctcactctt tggtaatcag gctgccgcaa
     4021 tatttatatc tgcctatatc ggcagttcta ttgtcggtaa ctcacttggg gcatggctgc
     4081 ttaccggcat ccggttcaat atcagaaaac tgattcaaat gcctcccgtc attgccctta
     4141 ttattggctg cttattgtta ccactgggta aacaaatccg cgagctggat ataatttatt
     4201 ccctcgctaa atttctgatg agctttcttg gcatggctat tcttggtatg tggctggcta
     4261 aaagcagaat tacactgacc gacatcagaa acaccattca gaccttcctg aaaaaagctg
     4321 cggcttattt ctgcatcctc tccgccctgc ttggccttgc ctgtctgtgc caacagaacc
     4381 ttattctgga aaatagtgcc gcactttact tattttgcct gcttcctccg gcagccaata
     4441 tcattgttct tgaaacccac tatctgggta ccggttactc cgttaaGAAT TCTACAgcgg
     4501 ccgcgattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4561 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4621 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4681 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg
     4741 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4801 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4861 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4921 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4981 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5041 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5101 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5161 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5221 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5281 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5341 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5401 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5461 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5581 gtgccacctg tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5641 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5701 ttgcaaac
//