LOCUS       sequence-66260-e        9404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA4.BDD FVIII.H4(R484H).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9404)
  TITLE     Peters lab - FVIII constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9404)
  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: 
            "Unpublished"
COMMENT     Sequence Label: BDD FVIII H4
FEATURES             Location/Qualifiers
     source          1..9404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4461..4490
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4506..4523
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4546..4563)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4552..4776
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4822..5250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4909..4928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5119..5140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5259..5279)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5264..5594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5445..5580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5507..5526
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5642..5689
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5708..6082
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6212..6333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6382..6398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6382..6398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6395..6417)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6406..6422
                     /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(6430..6436,6437..6454,6455..6460))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6475..6496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6613..6630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6784..7372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6864..6883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7543..8334,8335..8403))
                     /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     8166..8185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8404..8508)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8583..8602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8774..9153
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9154..9357
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9308..9328
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(9402..9404,1..17)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(9402..9404,1..16)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tacgactcac tatagggaga cccaagctgg ctagttaagc ttcgcgacgt acggccgcca
       61 ccatgcaaat agagctctcc acctgcttct ttctgtgcct tttgcgattc tgctttagtg
      121 ccaccagaag atactacctg ggtgcagtgg aactgtcatg ggactatatg caaagtgatc
      181 tcggtgagct gcctgtggac gcaagatttc ctcctagagt gccaaaatct tttccattca
      241 acacctcagt cgtgtacaaa aagactctgt ttgtagaatt cacggatcac cttttcaaca
      301 tcgctaagcc aaggccaccc tggatgggtc tgctaggtcc taccatccag gctgaggttt
      361 atgatacagt ggtcattaca cttaagaaca tggcttccca tcctgtcagt cttcatgctg
      421 ttggtgtatc ctactggaaa gcttctgagg gagctgaata tgatgatcag accagtcaaa
      481 gggagaaaga agatgataaa gtcttccctg gtggaagcca tacatatgtc tggcaggtcc
      541 tgaaagagaa tggtccaatg gcctctgacc cactgtgcct tacctactca tatctttctc
      601 atgtggacct ggtaaaagac ttgaattcag gcctcattgg agccctacta gtatgtagag
      661 aagggagtct ggccaaggaa aagacacaga ccttgcacaa atttatacta ctttttgctg
      721 tatttgatga agggaaaagt tggcactcag aaacaaagaa ctccttgatg caggataggg
      781 atgctgcatc tgctcgggcc tggcctaaaa tgcacacagt caatggttat gtaaacaggt
      841 ctctgccagg tctgattgga tgccacagga aatcagtcta ttggcatgtg attggaatgg
      901 gcaccactcc tgaagtgcac tcaatattcc tcgaaggtca cacatttctt gtgaggaacc
      961 atcgccaggc gtccttggaa atctcgccaa taactttcct tactgctcaa acactcttga
     1021 tggaccttgg acagtttcta ctgttttgtc atatctcttc ccaccaacat gatggcatgg
     1081 aagcttatgt caaagtagac agctgtccag aggaacccca actacgaatg aaaaataatg
     1141 aagaagcgga agactatgat gatgatctta ctgattctga aatggatgtg gtcaggtttg
     1201 atgatgacaa ctctccttcc tttatccaaa ttcgctcagt tgccaagaag catcctaaaa
     1261 cttgggtaca ttacattgct gctgaagagg aggactggga ctatgctccc ttagtcctcg
     1321 cccccgatga cagaagttat aaaagtcaat atttgaacaa tggccctcag cggattggta
     1381 ggaagtacaa aaaagtccga tttatggcat acacagatga aacctttaag actcgtgaag
     1441 ctattcagca tgaatcagga atcttgggac ctttacttta tggggaagtt ggagacacac
     1501 tgttgattat atttaagaat caagcaagca gaccatataa catctaccct cacggaatca
     1561 ctgatgtcca tcctttgtat tcaaggagat taccaaaagg tgtaaaacat ttgaaggatt
     1621 ttccaattct gccaggagaa atattcaaat ataaatggac agtgactgta gaagatgggc
     1681 caactaaatc agatcctcgg tgcctgaccc gctattactc tagtttcgtt aatatggaga
     1741 gagatctagc ttcaggactc attggccctc tcctcatctg ctacaaagaa tctgtagatc
     1801 aaagaggaaa ccagataatg tcagacaaga ggaatgtcat cctgttttct gtatttgatg
     1861 agaaccgaag ctggtacctc acagagaata tacaacgctt tctccccaat ccagctggag
     1921 tgcagcttga ggatccagag ttccaagcct ccaacatcat gcacagcatc aatggctatg
     1981 tttttgatag tttgcagttg tcagtttgtt tgcatgaggt ggcatactgg tacattctaa
     2041 gcattggagc acagactgac ttcctttctg tcttcttctc tggatatacc ttcaaacaca
     2101 aaatggtcta tgaagacaca ctcaccctat tcccattctc aggagaaact gtcttcatgt
     2161 cgatggaaaa cccaggtcta tggattctgg ggtgccacaa ctcagacttt cggaacagag
     2221 gcatgaccgc cttactgaag gtttctagtt gtgacaagaa cactggtgat tattacgagg
     2281 acagttatga agatatttca gcatacttgc tgagtaaaaa caatgccatt gaaccaagaa
     2341 gcttctctca aaacccacca gtcttgaaac gccatcaacg ggaaataact cgtactactc
     2401 ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa atgaagaagg
     2461 aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt caaaagaaaa
     2521 cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg agtagctccc
     2581 cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag aaagttgttt
     2641 tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa ctaaatgaac
     2701 atttgggact cctggggcca tatataagag cagaagttga agataatatc atggtaactt
     2761 tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct tatgaggaag
     2821 atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa accaaaactt
     2881 acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac tgcaaagcct
     2941 gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg attggacccc
     3001 ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg acagtacagg
     3061 aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc actgaaaata
     3121 tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact tttaaagaga
     3181 attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc ttagtaatgg
     3241 ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa aacatccatt
     3301 ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat aaaatggcac
     3361 tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc aaagctggaa
     3421 tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc acactttttc
     3481 tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac attagagatt
     3541 ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc agacttcatt
     3601 attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc aaggtggatc
     3661 tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag aagttctcca
     3721 gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag tggcagactt
     3781 atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat tcatctggga
     3841 taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg cacccaactc
     3901 attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta aatagttgca
     3961 gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact gcttcatcct
     4021 actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac ctccaaggga
     4081 ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa gtggacttcc
     4141 agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg cttaccagca
     4201 tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg actctctttt
     4261 ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca cctgtggtga
     4321 actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag agttgggtgc
     4381 accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc tactgagcgg
     4441 ccgctcgagg tcacccattc gaacaaaaac tcatctcaga agaggatctg aatatgcata
     4501 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     4561 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4621 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4681 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4741 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4801 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4861 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4921 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     4981 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5041 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5101 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5161 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     5221 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     5281 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     5341 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5401 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5461 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5521 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5581 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     5641 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     5701 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     5761 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     5821 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     5881 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     5941 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     6001 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     6061 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     6121 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6181 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6361 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6961 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8581 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8641 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8701 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     8761 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     8821 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     8881 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     8941 acgccaatag ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac
     9001 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     9061 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     9121 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     9181 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     9241 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     9301 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     9361 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaa
//
