LOCUS       pET-28a_PA4114_Cterm_Hi 5740 bp ds-DNA     circular     20-JAN-2023
DEFINITION  synthetic DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name: PA4114 Cterm
            His.txt NEBuilder-generated oligos (UPPERCASE = gene-specific,
            lowercase = overlap) PA4114_fwd:
            taagaaggagatataccatgATTGAACTGCGTACCATTACACGTG PA4114_fwd 3'Tm: 67.2
            3'Ta: 68.2 PA4114_rev: tggtggtggtggtgctcgagTGCCGGAACACCATGGGC
            PA4114_rev 3'Tm: 72.0  3'Ta: 68.2 pET-28a Cterm_His_fwd:
            CTCGAGCACCACCACCAC pET-28a Cterm_His_fwd 3'Tm: 68.9  3'Ta: 65.4
            pET-28a Cterm_His_rev: CATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAG
            pET-28a Cterm_His_rev 3'Tm: 64.4  3'Ta: 65.4
FEATURES             Location/Qualifiers
     primer_bind     2515..2534
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    2534..2556
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     protein_bind    2538..2554
                     /label="lac operator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(2562..2604)
                     /label="pET-28a Cterm_His_rev"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="gene-specific Tm: 64.4 Ta: 65.4"
                     /note="gene-specific primer: CATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAG"
     primer_bind     complement(2576..2604)
                     /label="pET28a Cterm Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     2585..2629
                     /label="PA4114_fwd"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="gene-specific Tm: 67.2 Ta: 68.2"
                     /note="overlap: taagaaggagatataccatg"
                     /note="gene-specific primer: ATTGAACTGCGTACCATTACACGTG"
     CDS             2602..3135
                     /label="PA4114_Cterm His"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MIELRTITRDDWETCIDLKVARHQAHFVASNLYSLAQSRFLPGFRAVGIHHGGRMVGFALYGPDADDGHYWLYRLMIDRRYQGKGHGRAALQRIIASVRRTRRRSDRLMVAYAVDNDVARQLYRDAGFVEQGLAPWGETMACLMLPEPSHRSGKVAGAPGGRAAHGVPALEHHHHHH*"
     gene            2605..3108
                     /label="PA4114"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(3091..3128)
                     /label="PA4114_rev"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="gene-specific Tm: 72.0 Ta: 68.2"
                     /note="overlap: tggtggtggtggtgctcgag"
                     /note="gene-specific primer: TGCCGGAACACCATGGGC"
     gene            3109..2604
                     /label="pET-28a Cterm_His"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     3109..3126
                     /label="pET-28a Cterm For"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3109..3126
                     /label="pET-28a Cterm_His_fwd"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="gene-specific Tm: 68.9 Ta: 65.4"
                     /note="gene-specific primer: CTCGAGCACCACCACCAC"
     primer_bind     3109..3129
                     /label="pET-28a For"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             3115..3132
                     /label="6x His Tag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             3115..3132
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      3283..3738
                     /label="M13 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      3300..3606
                     /label="F1 ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(3834..4646)
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      4692..5374
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5442..5462
                     /label="Seq Primer for Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(5541..5561)
                     /label="2,900 R"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
       61 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
      121 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
      181 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
      241 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
      301 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
      361 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
      421 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
      481 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
      541 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
      601 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
      661 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
      721 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
      781 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
      841 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
      901 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
      961 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     1021 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     1141 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     1201 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     1261 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     1321 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     1381 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     1441 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     1501 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     1561 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     1621 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     1681 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     1741 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     1801 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     1861 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     1921 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1981 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     2041 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     2101 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     2161 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     2221 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     2281 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     2341 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     2401 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     2461 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     2521 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     2581 actttaagaa ggagatatac catgattgaa ctgcgtacca ttacacgtga tgattgggaa
     2641 acctgtattg atctgaaagt tgcacgtcat caggcacatt ttgttgcaag caatctgtat
     2701 agcctggcac agagccgttt tctgcctggt tttcgtgcag ttggtattca tcatggtggt
     2761 cgtatggttg gttttgcact gtatggtccg gatgcagatg atggtcatta ttggctgtat
     2821 cgtctgatga ttgatcgtcg ctatcaaggt aaaggtcatg gtcgtgcagc actgcagcgt
     2881 attattgcaa gcgttcgtcg tacccgtcgt cgtagcgatc gcctgatggt tgcatatgcc
     2941 gttgataatg atgttgcccg tcagctgtac cgtgatgcag gttttgttga acaaggtctg
     3001 gcaccgtggg gtgaaaccat ggcatgtctg atgctgccgg aaccgagcca tcgtagcggt
     3061 aaagttgccg gtgcaccagg tggtcgcgca gcccatggtg ttccggcact cgagcaccac
     3121 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     3181 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     3241 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     3301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3721 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3781 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     3841 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     3901 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     3961 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     4021 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     4081 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     4141 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     4201 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     4261 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     4321 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     4381 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     4441 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     4501 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     4561 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     4621 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     4681 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4741 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4801 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4861 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4921 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4981 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5041 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5101 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5161 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5221 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5281 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5341 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5401 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5461 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5521 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5581 ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5641 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     5701 cacccgctga cgcgccctga cgggcttgtc tgctcccggc
//