LOCUS       sequence-408574-        8701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8701)
  AUTHORS   Du X, She E, Gelbart T, Truksa J, Lee P, Xia Y, Khovananth K, Mudd 
            S, Mann N, Moresco EM, Beutler E, Beutler B
  TITLE     The serine protease TMPRSS6 is required to sense iron deficiency.
  JOURNAL   Science. 2008 May 23. 320(5879):1088-92.
  PUBMED    18451267
REFERENCE   2  (bases 1 to 8701)
  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: "Science. 
            2008 May 23. 320(5879):1088-92."
COMMENT     Sequence Label: p3xFLAG-Tmprss6
FEATURES             Location/Qualifiers
     source          1..8701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2200..2209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3384..3449
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3460..4082
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4106..4126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4124..4421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4475..5269
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4529..4548)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5139..5158
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5921..6055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5958..5977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6012..6031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6060..6079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6061..6079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6093..6109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6093..6109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6106..6128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6117..6133
                     /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(6141..6147,6148..6165,6166..6171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6186..6207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6324..6341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6495..7083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6575..6594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7254..8045,8046..8114))
                     /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     7877..7896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8115..8219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8246..8674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8333..8352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8543..8564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaagctta agatgccgag atgtttccag ctcccctgtt
      961 ctaccaggat gcccaccacc gaggtccccc aagcggctga tggtcagggc gatgcgggtg
     1021 atggagagga agctgctgag ccagagggga agttcaagcc cccaaaaaac accaagagaa
     1081 aaaaccggga ctacgtccgc ttcacgccac tgttgctggt cttggctgcg ctggtctcag
     1141 caggggtcat gctttggtat ttcctagggt acaaagcgga agtgaccgta agccaggtgt
     1201 actctggcag cctccgggtg ctcaaccgtc atttctccca ggacctgggc cgacgggagt
     1261 ctattgcttt ccgcagtgaa tctgccaaag cccagaagat gctccaagaa ctggttgcca
     1321 gcacccgcct gggtacttac tacaactcta gttctgtcta ctcctttggg gagggacccc
     1381 tcacctgctt cttctggttt atccttgaca tccctgagta ccagcgactg accctgagcc
     1441 ctgaagtagt gcgcgagctc ctggtggatg agctactgtc caacagctca accctggctt
     1501 cctataagac cgaatatgag gtggacccgg aaggcctggt gatcctggaa gccagtgtga
     1561 acgacatagt cgtactgaat tccacgctgg gctgttatcg ctacagctat gtgaacccag
     1621 gccaggtcct cccattgaag gggcctgacc agcagaccac aagctgcctg tggcatctgc
     1681 aagggcccga agacctcatg atcaaagtgc ggctggagtg gacccgggtc gattgcagag
     1741 acagggtggc gatgtacgac gcagctgggc ccctggagaa gagacttatc acctcggtct
     1801 atgggtgcag ccgccaggaa cctgtgatgg aggtgctggc atcgggctcc gtcatggccg
     1861 tggtgtggaa aaagggcatg catagctact atgacccttt cctgctctca gtgaagtctg
     1921 tggccttcca ggactgccag gtgaacctga cactggaggg ccggctggac acacagggct
     1981 tcctccgtac accctactac cccagttact actctcccag tacccactgc tcctggcatc
     2041 tcacggtacc ctctctggac tacggcttgg cgctctggtt cgatgcctac gcactgagga
     2101 ggcagaagta caaccgactg tgtactcagg gccagtggat gatccagaac aggaggctgt
     2161 gtggcttccg taccctgcag ccatatgctg agaggatccc catggtggcc tcagatggtg
     2221 tcaccatcaa cttcacctcc cagatctccc tcacaggccc gggtgtgcaa gtgtactaca
     2281 gcttgtacaa ccaatcagac ccctgccctg gtgagttcct ctgctctgtg aatggactgt
     2341 gtgtccctgc gtgtgacggg atcaaggact gccccaatgg cctggatgag agaaactgtg
     2401 tctgcagagc catgttccag tgccaagagg acagcacgtg catttcactg cctagagtct
     2461 gtgaccggca gcccgactgt ctcaatggca gtgacgaaga acagtgccaa gaaggagtgc
     2521 cctgtgggac attcactttc cagtgtgagg accggagctg tgtgaagaag cccaacccag
     2581 agtgtgacgg ccagtcagat tgcagagacg gctcagatga gcaacactgt gactgtggcc
     2641 tccagggcct ctccagccgt attgtgggcg ggaccgtgtc ctccgagggt gagtggccat
     2701 ggcaggccag cctccagatt cggggtcgac acatctgtgg gggggctctc atcgctgacc
     2761 gctgggtcat aacggccgcc cactgcttcc aggaggacag catggcctcc ccgaagctgt
     2821 ggaccgtgtt cctgggaaag atgcggcaga actcgcgctg gccaggcgag gtgtccttca
     2881 aggtgagccg tctgttcctg cacccgtacc acgaggagga cagccatgac tacgacgtgg
     2941 ccctgctgca gctcgaccac cccgtggtgt actcggccac tgtgcgcccc gtctgcctgc
     3001 ctgcccgctc ccacttcttt gagccaggcc agcactgctg gatcacaggc tggggagccc
     3061 agcgagaggg tggtccggtg agcaacaccc tgcagaaggt ggacgtacag ctggtccctc
     3121 aggacctctg cagtgaggcc taccgctacc aggtgtcccc acgcatgctc tgtgctggct
     3181 accgcaaggg caagaaagat gcctgccagg gtgactctgg aggcccactg gtttgcaggg
     3241 agcccagtgg ccgctggttc ctggcagggt tggttagctg gggcctgggc tgtggccgac
     3301 ccaatttctt tggcgtctac acccgtgtca cacgtgtgat caactggatc cagcaggtgc
     3361 tgacctctag aggatcccgg gctgactaca aagaccatga cggtgattat aaagatcatg
     3421 acatcgacta caaggatgac gatgacaagt agtgatcccg ggtggcatcc ctgtgacccc
     3481 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3541 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3601 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3661 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3721 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3781 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3841 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3901 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     3961 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     4021 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     4081 ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4201 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4261 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4321 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4381 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggaattga tcagcttggg
     4441 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4501 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4561 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4621 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4681 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4741 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4801 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4861 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4921 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4981 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5041 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5101 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5161 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5221 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5281 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5341 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5401 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccgggctcga tcccctcgcg
     5461 agttggttca gctgctgcct gaggctggac gacctcgcgg agttctaccg gcagtgcaaa
     5521 tccgtcggca tccaggaaac cagcagcggc tatccgcgca tccatgcccc cgaactgcag
     5581 gagtggggag gcacgatggc cgctttggtc gacccggacg ggacgctcct gcgcctgata
     5641 cagaacgaat tgcttgcagg catctcatga gtgtgtcttc ccgttttccg cctgaggtca
     5701 ctgcgtggat ggagcgctgg cgcctgctgc gcgacggcga gctgctcacc acccactcgc
     5761 caagctggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgccgatcat
     5821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6001 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaattc
     6061 cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     6121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6901 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8221 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     8641 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     8701 t
//
