LOCUS       pET-28a_PA1377_Nterm_Hi 5827 bp ds-DNA     circular     29-APR-2023
DEFINITION  synthetic DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            pET-28a_PA1377_Nhis.txt NEBuilder-generated oligos (UPPERCASE =
            gene-specific, lowercase = overlap) PA1377_fwd:
            tggtgccgcgcggcagccatATGACCGCAGAAAGCCCG PA1377_fwd 3'Tm: 68.6  3'Ta:
            65.6 PA1377_rev: tggtggtggtggtgctcgagTTATTCACCAACACGACCTTCC
            PA1377_rev 3'Tm: 64.6  3'Ta: 65.6 pET-28a_Nhis_fwd:
            CTCGAGCACCACCACCACCACC pET-28a_Nhis_fwd 3'Tm: 75.5  3'Ta: 72.0
            pET-28a_Nhis_rev: ATGGCTGCCGCGCGGCAC pET-28a_Nhis_rev 3'Tm: 79.0
            3'Ta: 72.0
FEATURES             Location/Qualifiers
     primer_bind     450..469
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    469..491
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     protein_bind    473..489
                     /label="lac operator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(511..539)
                     /label="pET28a Cterm Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     577..614
                     /label="PA1377_fwd"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene-specific Tm: 68.6 Ta: 65.6"
                     /note="overlap: tggtgccgcgcggcagccat"
                     /note="gene-specific primer: ATGACCGCAGAAAGCCCG"
     primer_bind     complement(579..596)
                     /label="pET-28 Rev"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     complement(579..596)
                     /label="pET-28a_Nhis_rev"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="gene-specific Tm: 79.0 Ta: 72.0"
                     /note="gene-specific primer: ATGGCTGCCGCGCGGCAC"
     gene            597..1130
                     /label="PA1377"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             597..1130
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MTAESPTIRLERYSERHVEGLTALYNDPAVARQVLQMPYQSVEQRRKRLHDSADDDRLLILVALHQGDVIGSASLEQHPRIRRSHSGSIGMGVAVAWQGKGVGSRLLGELLDIADNWMNLRRVELTVYTDNAPALALYRKFGFETEGEMRDYAVRDGRFVDVYSMARLRRVEGRVGE*"
     primer_bind     complement(1109..1150)
                     /label="PA1377_rev"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="gene-specific Tm: 64.6 Ta: 65.6"
                     /note="overlap: tggtggtggtggtgctcgag"
                     /note="gene-specific primer: TTATTCACCAACACGACCTTCC"
     gene            1131..596
                     /label="pET-28a_Nhis"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     1131..1148
                     /label="pET-28a Cterm For"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     1131..1151
                     /label="pET-28a For"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer_bind     1131..1152
                     /label="pET-28a_Nhis_fwd"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="gene-specific Tm: 75.5 Ta: 72.0"
                     /note="gene-specific primer: CTCGAGCACCACCACCACCACC"
     CDS             1137..1154
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             1137..1154
                     /label="6x His Tag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      1305..1760
                     /label="M13 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      1322..1628
                     /label="F1 ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(1856..2668)
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      2714..3396
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3464..3484
                     /label="Seq Primer for Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(3563..3583)
                     /label="2,900 R"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
       61 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
      121 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
      181 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
      241 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
      301 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
      361 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
      421 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
      481 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
      541 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatga
      601 ccgcagaaag cccgaccatt cgtctggaac gttatagcga acgtcatgtt gaaggtctga
      661 ccgcactgta taatgatccg gcagttgcac gtcaggttct gcagatgccg tatcagagcg
      721 ttgaacagcg tcgtaaacgt ctgcatgata gcgcagatga tgatcgtctg ctgattctgg
      781 ttgcactgca tcagggtgat gttattggta gcgcaagcct ggaacagcat ccgcgtattc
      841 gtcgtagcca tagcggtagc attggtatgg gtgttgcagt tgcatggcaa ggtaaaggtg
      901 ttggtagccg tctgctgggt gaactgctgg atattgcaga taattggatg aatctgcgtc
      961 gtgttgaact gaccgtttat accgataatg caccggcact ggcactgtat cgtaaatttg
     1021 gttttgaaac cgaaggtgag atgcgtgatt atgcagttcg tgatggtcgt tttgttgatg
     1081 tttatagcat ggcacgtctg cgtcgcgtgg aaggtcgtgt tggtgaataa ctcgagcacc
     1141 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1201 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1261 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     1321 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1381 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1441 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1501 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1561 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1621 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1681 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1741 aatattaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     1861 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1921 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1981 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     2041 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     2101 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     2161 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2221 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2281 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2341 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2401 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2461 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2521 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2581 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     2641 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     2701 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3481 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3541 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3601 atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3661 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3721 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3781 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3841 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3901 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3961 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4021 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4081 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4141 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4201 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4261 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     4321 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     4381 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4441 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     4501 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4561 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4621 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4681 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4741 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4921 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4981 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     5041 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     5101 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5161 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     5221 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5281 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5341 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5401 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5461 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     5521 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5581 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5641 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5701 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5761 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5821 ccacgcg
//