LOCUS       pET-28a_PA4114_Nterm_Hi 5803 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 Nterm
            His.txt NEBuilder-generated oligos (UPPERCASE = gene-specific,
            lowercase = overlap) PA4114_fwd:
            tggtgccgcgcggcagccatATGATTGAACTGCGTACC PA4114_fwd 3'Tm: 59.1  3'Ta:
            60.1 PA4114_rev: tggtggtggtggtgctcgagTTATGCCGGAACACCATG PA4114_rev
            3'Tm: 62.3  3'Ta: 60.1 pET-28a Nterm_His_fwd:
            CTCGAGCACCACCACCACCACC pET-28a Nterm_His_fwd 3'Tm: 75.5  3'Ta: 72.0
            pET-28a Nterm_His_rev: ATGGCTGCCGCGCGGCAC pET-28a Nterm_His_rev
            3'Tm: 79.0  3'Ta: 72.0
FEATURES             Location/Qualifiers
     primer_bind     138..157
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    157..179
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     protein_bind    161..177
                     /label="lac operator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(199..227)
                     /label="pET28a Cterm Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             225..794
                     /label="PA4114 Nterm_His"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MGSSHHHHHHSSGLVPRGSHMIELRTITRDDWETCIDLKVARHQAHFVASNLYSLAQSRFLPGFRAVGIHHGGRMVGFALYGPDADDGHYWLYRLMIDRRYQGKGHGRAALQRIIASVRRTRRRSDRLMVAYAVDNDVARQLYRDAGFVEQGLAPWGETMACLMLPEPSHRSGKVAGAPGGRAAHGVPA*"
     primer_bind     265..302
                     /label="PA4114_fwd"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="gene-specific Tm: 59.1 Ta: 60.1"
                     /note="overlap: tggtgccgcgcggcagccat"
                     /note="gene-specific primer: ATGATTGAACTGCGTACC"
     primer_bind     complement(267..284)
                     /label="pET-28a Nterm_His_rev"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="gene-specific Tm: 79.0 Ta: 72.0"
                     /note="gene-specific primer: ATGGCTGCCGCGCGGCAC"
     primer_bind     complement(267..284)
                     /label="pET-28 Rev"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     gene            285..794
                     /label="PA4114"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(777..814)
                     /label="PA4114_rev"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="gene-specific Tm: 62.3 Ta: 60.1"
                     /note="overlap: tggtggtggtggtgctcgag"
                     /note="gene-specific primer: TTATGCCGGAACACCATG"
     gene            795..284
                     /label="pET-28a Nterm_His"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     795..812
                     /label="pET-28a Cterm For"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     795..815
                     /label="pET-28a For"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer_bind     795..816
                     /label="pET-28a Nterm_His_fwd"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="gene-specific Tm: 75.5 Ta: 72.0"
                     /note="gene-specific primer: CTCGAGCACCACCACCACCACC"
     CDS             801..818
                     /label="6x His Tag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             801..818
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      969..1424
                     /label="M13 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      986..1292
                     /label="F1 ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(1520..2332)
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      2378..3060
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3128..3148
                     /label="Seq Primer for Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(3227..3247)
                     /label="2,900 R"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
       61 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
      121 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
      181 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc
      241 atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatgatt gaactgcgta
      301 ccattacacg tgatgattgg gaaacctgta ttgatctgaa agttgcacgt catcaggcac
      361 attttgttgc aagcaatctg tatagcctgg cacagagccg ttttctgcct ggttttcgtg
      421 cagttggtat tcatcatggt ggtcgtatgg ttggttttgc actgtatggt ccggatgcag
      481 atgatggtca ttattggctg tatcgtctga tgattgatcg tcgctatcaa ggtaaaggtc
      541 atggtcgtgc agcactgcag cgtattattg caagcgttcg tcgtacccgt cgtcgtagcg
      601 atcgcctgat ggttgcatat gccgttgata atgatgttgc ccgtcagctg taccgtgatg
      661 caggttttgt tgaacaaggt ctggcaccgt ggggtgaaac catggcatgt ctgatgctgc
      721 cggaaccgag ccatcgtagc ggtaaagttg ccggtgcacc aggtggtcgc gcagcccatg
      781 gtgttccggc ataactcgag caccaccacc accaccactg agatccggct gctaacaaag
      841 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      901 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
      961 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1021 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1081 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1141 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1201 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1261 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1321 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1381 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg
     1441 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1501 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1561 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1621 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1681 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1741 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     1801 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1861 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     1921 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1981 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2041 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2101 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2161 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2221 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2281 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     2341 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     2401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2581 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2821 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3121 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3181 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3241 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     3301 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     3361 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3421 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3481 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     3541 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     3601 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     3661 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     3721 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     3781 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     3841 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     3901 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     3961 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     4021 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     4081 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     4141 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     4201 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     4261 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     4321 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     4381 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     4441 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     4621 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     4681 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     4741 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     4801 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     4861 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     4921 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     4981 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     5041 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5101 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     5161 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     5221 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     5281 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     5341 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     5401 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     5461 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     5521 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     5581 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     5641 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     5701 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     5761 gccacggggc ctgccaccat acccacgccg aaacaagcgc tca
//