LOCUS       sequence-129388-        8074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-His tagged nonglycosylated human serum transferrin 
            unable to bind iron in either the N-lobe or the C-lobeN-lobe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8074)
  AUTHORS   Mason AB, Halbrooks PJ, Larouche JR, Briggs SK, Moffett ML, Ramsey 
            JE, Connolly SA, Smith VC, MacGillivray RT
  TITLE     Expression, purification, and characterization of authentic 
            monoferric and apo-human serum transferrins.
  JOURNAL   Protein Expr Purif. 2004 Aug;36(2):318-26.
  PUBMED    15249056
REFERENCE   2  (bases 1 to 8074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr Purif."; date: "2004-08"; volume: "36(2)"; pages: "318-26"
COMMENT     Sequence Label: pNUT N6His Y95F/Y188F/Y426F/Y517F hTFNG hTFNG
FEATURES             Location/Qualifiers
     source          1..8074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             70..87
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             88..99
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2311..2933
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2943..2959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2943..2959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2956..2978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2967..2983
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2991..2997,2998..3015,3016..3021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3036..3057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3174..3191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3345..3933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3425..3444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4104..4895,4896..4964))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4727..4746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4965..5069)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5137..5155
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5193..5215)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5315..5334
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5528..5550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5542..5559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5543..5559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5780..5799
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6526..7089)
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDRPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     promoter        complement(7117..7474)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7131..7266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7283..7474
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7431..7451
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 atgaggctcg ccgtgggagc cctgctggtc tgcgccgtcc tagggctgtg tctggctgtc
       61 cctgataaac atcatcatca tcatcatatc gagggaaggg tccctgataa aactgtgaga
      121 tggtgtgcag tgtcggagca tgaggccact aagtgccaga gtttccgcga ccatatgaaa
      181 agcgtcattc catccgatgg tcccagtgtt gcttgtgtga agaaagcctc ctaccttgat
      241 tgcatcaggg ccattgcggc aaacgaagcg gatgctgtga cactggatgc aggtttggtg
      301 tatgatgctt acctggctcc caataacctg aagcctgtgg tggcagagtt ctatgggtca
      361 aaagaggatc cacagacttt cttttatgct gttgctgtgg tgaagaagga tagtggcttc
      421 cagatgaacc agcttcgagg caagaagtcc tgccacacgg gtctaggcag gtccgctggg
      481 tggaacatcc ccataggctt actttactgt gacttacctg agccacgtaa acctcttgag
      541 aaagcagtgg ccaatttctt ctcgggcagc tgtgcccctt gtgcggatgg gacggacttc
      601 ccccagctgt gtcaactgtg tccagggtgt ggctgctcca cccttaacca atacttcggc
      661 ttctcgggag ccttcaagtg tctgaaggat ggtgctgggg atgtggcctt tgtcaagcac
      721 tcgactatat ttgagaactt ggcaaacaag gctgacaggg accagtatga gctgctttgc
      781 ctggacaaca cccggaagcc ggtagatgaa tacaaggact gccacttggc ccaggtccct
      841 tctcataccg tcgtggcccg aagtatgggc ggcaaggagg acttgatctg ggagcttctc
      901 aaccaggccc aggaacattt tggcaaagac aaatcaaaag aattccaact attcagctct
      961 cctcatggga aggacctgct gtttaaggac tctgcccacg ggtttttaaa agtccccccc
     1021 aggatggatg ccaagatgta cctgggctat gagtatgtca ctgccatccg gaatctacgg
     1081 gaaggcacat gcccagaagc cccaacagat gaatgcaagc ctgtgaagtg gtgtgcgctg
     1141 agccaccacg agaggctcaa gtgtgatgag tggagtgtta acagtgtagg gaaaatagag
     1201 tgtgtatcag cagagaccac cgaagactgc atcgccaaga tcatgaatgg agaagctgat
     1261 gccatgagct tggatggagg gtttgtctac atagcgggca agtgtggtct ggtgcctgtc
     1321 ttggcagaaa actacgataa gagcgataat tgtgaggata caccagaggc agggtttttc
     1381 gctgtagcag tggtgaagaa atcagcttct gacctcacct gggacaatct gaaaggcaag
     1441 aagtcctgcc atacggcagt tggcagaacc gctggctgga acatccccat gggcctgctc
     1501 tacaataaga tcaaccactg cagatttgat gaatttttca gtgaaggttg tgcccctggg
     1561 tctaagaaag actccagtct ctgtaagctg tgtatgggct caggcctaaa cctgtgtgaa
     1621 cccaacaaca aagagggata ctacggcttc acaggcgctt tcaggtgtct ggttgagaag
     1681 ggagatgtgg cctttgtgaa acaccagact gtcccacaga acactggggg aaaaaaccct
     1741 gatccatggg ctaagaatct gaatgaaaaa gactatgagt tgctgtgcct tgatggtacc
     1801 aggaaacctg tggaggagta tgcgaactgc cacctggcca gagccccgaa tcacgctgtg
     1861 gtcacacgga aagataagga agcttgcgtc cacaagatat tacgtcaaca gcagcaccta
     1921 tttggaagcg acgtaactga ctgctcgggc aacttttgtt tgttccggtc ggaaaccaag
     1981 gaccttctgt tcagagatga cacagtatgt ttggccaaac ttcatgacag aaacacatat
     2041 gaaaaatact taggagaaga atatgtcaag gctgttggta acctgagaaa atgctccacc
     2101 tcatcactcc tggaagcctg cactttccgt agaccttaaa atctcagagg tagggctgcc
     2161 accaaggtga agatgggaac gcagatgatc catgagtttg ccctggtttc actggcccaa
     2221 gtggtttgtg ctaaccacgt ctgtcttcac agctctgtgt tgccatgtgt gctgaacaaa
     2281 aaataaaaat tattattgat tttatatttc gggtggcatc cctgtgaccc ctccccagtg
     2341 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2401 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2461 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2521 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     2581 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     2641 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     2701 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     2761 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     2821 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     2881 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attcgtaatc
     2941 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3061 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3121 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3181 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3241 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3301 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3361 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3421 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3481 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3541 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3601 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3661 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3721 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3781 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3841 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3901 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3961 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4021 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4081 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4141 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4201 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4261 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4321 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4381 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4441 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4501 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4561 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4621 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4681 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4741 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4801 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4861 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4921 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5101 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5161 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5221 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5281 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5341 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5401 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5461 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5521 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttgc atgcctgcag
     5581 gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5641 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5701 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5761 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5821 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctctgacgga aggaaagaag
     5881 tcagaaggca aaaacgagag taactccaca gtagctccaa attctttata agggtcaatg
     5941 tccatgcccc aaagccaccc aaggcacagc ttggaggctt gaacagtggg acatgtacaa
     6001 gagatgatta ggcagaggtg aaaaagttgc atggtgctgg tgcgcagacc aatttatgcc
     6061 tacagcctcc taatacaaag acctttaacc taatctcctc ccccagctcc tcccagtcct
     6121 taaacacaca gtctttgaag taggcctcaa ggtcggtcgt tgacattgct gggagtccaa
     6181 gagtcctctt atgtaagacc ttgggcagga tctgatgggc gttcacggtg gtctccatgc
     6241 aacgtgcaga ggtgaagcga agtgcacacg gaccggcaga tgagaaggca cagacgggga
     6301 gaccgcgtaa agagaggtgc gccccgtggt cggctggaac ggcagacgga gaaggggacg
     6361 agagagtccc aagcggcccc gcgaggggtc gtccgcggga ttcagcgccg acgggacgta
     6421 aacaaaggac gtcccgcgaa ggatctaaag ccagcaaaag tcccatggtc ttataaaaat
     6481 gcatagcttt aggaggggag cagagaactt gaaagcatct tcctgttagt ctttcttctc
     6541 gtagacttca aacttatact tgatgccttt ttcctcctgg acctcagaga ggacgcctgg
     6601 gtattctggg agaagtttat atttccccaa atcaatttct gggaaaaacg tgtcactttc
     6661 aaattcctgc atgatccttg tcacaaagag tctgaggtgg cctggttgat tcatggcttc
     6721 ctggtaaaca gaactgcctc cgactatcca aaccatgtct actttacttg ccaattccgg
     6781 ttgttcaata agtcttaagg catcatccaa acttttggca agaaaatgag ctcctcgtgg
     6841 tggttctttg agttctctac tgagaactat attaattctg tcctttaaag gtcgattctt
     6901 ctcaggaatg gagaaccagg ttttcctacc cataatcacc agattctgtt taccttccac
     6961 tgaagaggtt gtggtcattc tttggaagta cttgaactcg ttcctgagcg gaggccaggg
     7021 tcggtctccg ttcttgccaa tccccatatt ttgggacacg gcgacgatgc agttcaatgg
     7081 tcgaaccatg atggcaattc tagaatcgat aagctttttg caaaagccta ggcctccaaa
     7141 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     7201 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     7261 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     7321 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     7381 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     7441 acacacattc cacagctggt tctttccgcc tcagaaggta ccttggtttt taaaaccagc
     7501 ctggagtaga gcagatgggt taaggtgagt gacccctcag ccctggacat tcttagatga
     7561 gccccctcag gagtagagaa taatgttgag atgagttctg ttggctaaaa taatcaaggc
     7621 tagtctttat aaaactgtct cctcttctcc tagcttcgat ccagagagag acctgggcgg
     7681 agctggtcgc tgctcaggaa ctccaggaaa ggagaagctg aggttaccac gctgcgaatg
     7741 ggtttacgga gatagctggc tttccggggt gagttctcgt aaactccaga gcagcgatag
     7801 gccgtaatat cggggaaagc actataggga catgatgttc cacacgtcac atgggtcgtc
     7861 ctatccgagc cagtcgtgcc aaaggggcgg tcccgctgtg cacactggcg ctccagggag
     7921 ctctgcactc cgcccgaaaa gtgcgctcgg ctctgccaag gacgcggggc gcgtgactat
     7981 gcgtgggctg gagcaaccgc ctgctgggtg caaacccttt gcgcccggac tcgtccaacg
     8041 actataaaga gggcaggctg tcctctaagc gtca
//
