LOCUS       sequence-124401-        7952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-His tagged nonglycosylated human serum transferrin in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7952)
  AUTHORS   Mason AB, He QY, Halbrooks PJ, Everse SJ, Gumerov DR, Kaltashov IA, 
            Smith VC, Hewitt J, MacGillivray RT
  TITLE     Differential effect of a his tag at the N- and C-termini: functional
            studies with recombinant human serum transferrin.
  JOURNAL   Biochemistry. 2002 Jul 30;41(30):9448-54.
  PUBMED    12135367
REFERENCE   2  (bases 1 to 7952)
  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: 
            "Biochemistry."; date: "2002-07-30"; volume: "41(30)"; pages: 
            "9448-54"
COMMENT     Sequence Label: pNUT N6His hTFNG
FEATURES             Location/Qualifiers
     source          1..7952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..96
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             97..108
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2189..2811
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2388..2407)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2821..2837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2821..2837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2834..2856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2845..2861
                     /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(2869..2875,2876..2893,2894..2899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2914..2935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3052..3069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3303..3322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3982..4773,4774..4842))
                     /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     4605..4624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4843..4947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5015..5033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5071..5093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5193..5212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5406..5428
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5420..5437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5421..5437
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5658..5677
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6404..6967)
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDRPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     promoter        complement(6995..7352)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7009..7144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7161..7352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7309..7329
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cccgggaaga tgaggctcgc cgtgggagcc ctgctggtct gcgccgtcct agggctgtgt
       61 ctggctgtcc ctgataaaca tcatcatcat catcatatcg agggaagggt ccctgataaa
      121 actgtgagat ggtgtgcagt gtcggagcat gaggccacta agtgccagag tttccgcgac
      181 catatgaaaa gcgtcattcc atccgatggt cccagtgttg cttgtgtgaa gaaagcctcc
      241 taccttgatt gcatcagggc cattgcggca aacgaagcgg atgctgtgac actggatgca
      301 ggtttggtgt atgatgctta cctggctccc aataacctga agcctgtggt ggcagagttc
      361 tatgggtcaa aagaggatcc acagactttc tattatgctg ttgctgtggt gaagaaggat
      421 agtggcttcc agatgaacca gcttcgaggc aagaagtcct gccacacggg tctaggcagg
      481 tccgctgggt ggaacatccc cataggctta ctttactgtg acttacctga gccacgtaaa
      541 cctcttgaga aagcagtggc caatttcttc tcgggcagct gtgccccttg tgcggatggg
      601 acggacttcc cccagctgtg tcaactgtgt ccagggtgtg gctgctccac ccttaaccaa
      661 tacttcggct actcgggagc cttcaagtgt ctgaaggatg gtgctgggga tgtggccttt
      721 gtcaagcact cgactatatt tgagaacttg gcaaacaagg ctgacaggga ccagtatgag
      781 ctgctttgcc tggacaacac ccggaagccg gtagatgaat acaaggactg ccacttggcc
      841 caggtccctt ctcataccgt cgtggcccga agtatgggcg gcaaggagga cttgatctgg
      901 gagcttctca accaggccca ggaacatttt ggcaaagaca aatcaaaaga attccaacta
      961 ttcagctctc ctcatgggaa ggacctgctg tttaaggact ctgcccacgg gtttttaaaa
     1021 gtccccccca ggatggatgc caagatgtac ctgggctatg agtatgtcac tgccatccgg
     1081 aatctacggg aaggcacatg cccagaagcc ccaacagatg aatgcaagcc tgtgaagtgg
     1141 tgtgcgctga gccaccacga gaggctcaag tgtgatgagt ggagtgttaa cagtgtaggg
     1201 aaaatagagt gtgtatcagc agagaccacc gaagactgca tcgccaagat catgaatgga
     1261 gaagctgatg ccatgagctt ggatggaggg tttgtctaca tagcgggcaa gtgtggtctg
     1321 gtgcctgtct tggcagaaaa ctacgataag agcgataatt gtgaggatac accagaggca
     1381 gggtattttg ctgtagcagt ggtgaagaaa tcagcttctg acctcacctg ggacaatctg
     1441 aaaggcaaga agtcctgcca tacggcagtt ggcagaaccg ctggctggaa catccccatg
     1501 ggcctgctct acaataagat caaccactgc agatttgatg aatttttcag tgaaggttgt
     1561 gcccctgggt ctaagaaaga ctccagtctc tgtaagctgt gtatgggctc aggcctaaac
     1621 ctgtgtgaac ccaacaacaa agagggatac tacggctaca caggcgcttt caggtgtctg
     1681 gttgagaagg gagatgtggc ctttgtgaaa caccagactg tcccacagaa cactggggga
     1741 aaaaaccctg atccatgggc taagaatctg aatgaaaaag actatgagtt gctgtgcctt
     1801 gatggtacca ggaaacctgt ggaggagtat gcgaactgcc acctggccag agccccgaat
     1861 cacgctgtgg tcacacggaa agataaggaa gcttgcgtcc acaagatatt acgtcaacag
     1921 cagcacctat ttggaagcga cgtaactgac tgctcgggca acttttgttt gttccggtcg
     1981 gaaaccaagg accttctgtt cagagatgac acagtatgtt tggccaaact tcatgacaga
     2041 aacacatatg aaaaatactt aggagaagaa tatgtcaagg ctgttggtaa cctgagaaaa
     2101 tgctccacct catcactcct ggaagcctgc actttccgta gaccttaata gctcgagggc
     2161 ggccgctgat tgattgagtc gactctaggg gtggcatccc tgtgacccct ccccagtgcc
     2221 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     2281 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     2341 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     2401 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     2461 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     2521 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     2581 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     2641 aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt
     2701 ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct
     2761 tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tcgtaatcat
     2821 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2881 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2941 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3001 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4981 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5041 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5101 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5161 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5221 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5281 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5341 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5401 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcctgcaggt
     5461 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     5521 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     5581 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     5641 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     5701 ccggtgatgc cggccacgat gcgtccggcg tagaggatct ctgacggaag gaaagaagtc
     5761 agaaggcaaa aacgagagta actccacagt agctccaaat tctttataag ggtcaatgtc
     5821 catgccccaa agccacccaa ggcacagctt ggaggcttga acagtgggac atgtacaaga
     5881 gatgattagg cagaggtgaa aaagttgcat ggtgctggtg cgcagaccaa tttatgccta
     5941 cagcctccta atacaaagac ctttaaccta atctcctccc ccagctcctc ccagtcctta
     6001 aacacacagt ctttgaagta ggcctcaagg tcggtcgttg acattgctgg gagtccaaga
     6061 gtcctcttat gtaagacctt gggcaggatc tgatgggcgt tcacggtggt ctccatgcaa
     6121 cgtgcagagg tgaagcgaag tgcacacgga ccggcagatg agaaggcaca gacggggaga
     6181 ccgcgtaaag agaggtgcgc cccgtggtcg gctggaacgg cagacggaga aggggacgag
     6241 agagtcccaa gcggccccgc gaggggtcgt ccgcgggatt cagcgccgac gggacgtaaa
     6301 caaaggacgt cccgcgaagg atctaaagcc agcaaaagtc ccatggtctt ataaaaatgc
     6361 atagctttag gaggggagca gagaacttga aagcatcttc ctgttagtct ttcttctcgt
     6421 agacttcaaa cttatacttg atgccttttt cctcctggac ctcagagagg acgcctgggt
     6481 attctgggag aagtttatat ttccccaaat caatttctgg gaaaaacgtg tcactttcaa
     6541 attcctgcat gatccttgtc acaaagagtc tgaggtggcc tggttgattc atggcttcct
     6601 ggtaaacaga actgcctccg actatccaaa ccatgtctac tttacttgcc aattccggtt
     6661 gttcaataag tcttaaggca tcatccaaac ttttggcaag aaaatgagct cctcgtggtg
     6721 gttctttgag ttctctactg agaactatat taattctgtc ctttaaaggt cgattcttct
     6781 caggaatgga gaaccaggtt ttcctaccca taatcaccag attctgttta ccttccactg
     6841 aagaggttgt ggtcattctt tggaagtact tgaactcgtt cctgagcgga ggccagggtc
     6901 ggtctccgtt cttgccaatc cccatatttt gggacacggc gacgatgcag ttcaatggtc
     6961 gaaccatgat ggcaattcta gaatcgataa gctttttgca aaagcctagg cctccaaaaa
     7021 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     7081 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     7141 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     7201 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     7261 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     7321 acacattcca cagctggttc tttccgcctc agaaggtacc ttggttttta aaaccagcct
     7381 ggagtagagc agatgggtta aggtgagtga cccctcagcc ctggacattc ttagatgagc
     7441 cccctcagga gtagagaata atgttgagat gagttctgtt ggctaaaata atcaaggcta
     7501 gtctttataa aactgtctcc tcttctccta gcttcgatcc agagagagac ctgggcggag
     7561 ctggtcgctg ctcaggaact ccaggaaagg agaagctgag gttaccacgc tgcgaatggg
     7621 tttacggaga tagctggctt tccggggtga gttctcgtaa actccagagc agcgataggc
     7681 cgtaatatcg gggaaagcac tatagggaca tgatgttcca cacgtcacat gggtcgtcct
     7741 atccgagcca gtcgtgccaa aggggcggtc ccgctgtgca cactggcgct ccagggagct
     7801 ctgcactccg cccgaaaagt gcgctcggct ctgccaagga cgcggggcgc gtgactatgc
     7861 gtgggctgga gcaaccgcct gctgggtgca aaccctttgc gcccggactc gtccaacgac
     7921 tataaagagg gcaggctgtc ctctaagcgt ca
//
