LOCUS       sequence-263882-        9403 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 9403)
  AUTHORS   Peters RT, Toby G, Lu Q, Liu T, Kulman JD, Low SC, Bitonti AJ, 
            Pierce GF
  TITLE     Biochemical and functional characterization of a recombinant 
            monomeric Factor VIII-Fc fusion protein.
  JOURNAL   J Thromb Haemost. 2012 Dec 4. doi: 10.1111/jth.12076.
  PUBMED    23205847
REFERENCE   2  (bases 1 to 9403)
  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: "J Thromb 
            Haemost. 2012 Dec 4. doi: 10.1111/jth.12076."
COMMENT     Sequence Label: pCDNA4/BDD-FVIII
FEATURES             Location/Qualifiers
     source          1..9403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5092..5121
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5137..5154
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5177..5194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5183..5407
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5453..5881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5540..5559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5750..5771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5890..5910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5895..6224
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6075..6210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6137..6156
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6272..6319
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6338..6712
                     /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    6842..6963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6879..6898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6933..6952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7012..7028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7012..7028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7025..7047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7036..7052
                     /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(7060..7066,7067..7084,7085..7090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7105..7126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7243..7260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7414..8002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7494..7513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8173..8964,8965..9033))
                     /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     8796..8815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9034..9138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9213..9232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttcgcgacg tacggccgcc accatgcaaa tagagctctc cacctgcttc
      721 tttctgtgcc ttttgcgatt ctgctttagt gccaccagaa gatactacct gggtgcagtg
      781 gaactgtcat gggactatat gcaaagtgat ctcggtgagc tgcctgtgga cgcaagattt
      841 cctcctagag tgccaaaatc ttttccattc aacacctcag tcgtgtacaa aaagactctg
      901 tttgtagaat tcacggatca ccttttcaac atcgctaagc caaggccacc ctggatgggt
      961 ctgctaggtc ctaccatcca ggctgaggtt tatgatacag tggtcattac acttaagaac
     1021 atggcttccc atcctgtcag tcttcatgct gttggtgtat cctactggaa agcttctgag
     1081 ggagctgaat atgatgatca gaccagtcaa agggagaaag aagatgataa agtcttccct
     1141 ggtggaagcc atacatatgt ctggcaggtc ctgaaagaga atggtccaat ggcctctgac
     1201 ccactgtgcc ttacctactc atatctttct catgtggacc tggtaaaaga cttgaattca
     1261 ggcctcattg gagccctact agtatgtaga gaagggagtc tggccaagga aaagacacag
     1321 accttgcaca aatttatact actttttgct gtatttgatg aagggaaaag ttggcactca
     1381 gaaacaaaga actccttgat gcaggatagg gatgctgcat ctgctcgggc ctggcctaaa
     1441 atgcacacag tcaatggtta tgtaaacagg tctctgccag gtctgattgg atgccacagg
     1501 aaatcagtct attggcatgt gattggaatg ggcaccactc ctgaagtgca ctcaatattc
     1561 ctcgaaggtc acacatttct tgtgaggaac catcgccagg cgtccttgga aatctcgcca
     1621 ataactttcc ttactgctca aacactcttg atggaccttg gacagtttct actgttttgt
     1681 catatctctt cccaccaaca tgatggcatg gaagcttatg tcaaagtaga cagctgtcca
     1741 gaggaacccc aactacgaat gaaaaataat gaagaagcgg aagactatga tgatgatctt
     1801 actgattctg aaatggatgt ggtcaggttt gatgatgaca actctccttc ctttatccaa
     1861 attcgctcag ttgccaagaa gcatcctaaa acttgggtac attacattgc tgctgaagag
     1921 gaggactggg actatgctcc cttagtcctc gcccccgatg acagaagtta taaaagtcaa
     1981 tatttgaaca atggccctca gcggattggt aggaagtaca aaaaagtccg atttatggca
     2041 tacacagatg aaacctttaa gactcgtgaa gctattcagc atgaatcagg aatcttggga
     2101 cctttacttt atggggaagt tggagacaca ctgttgatta tatttaagaa tcaagcaagc
     2161 agaccatata acatctaccc tcacggaatc actgatgtcc gtcctttgta ttcaaggaga
     2221 ttaccaaaag gtgtaaaaca tttgaaggat tttccaattc tgccaggaga aatattcaaa
     2281 tataaatgga cagtgactgt agaagatggg ccaactaaat cagatcctcg gtgcctgacc
     2341 cgctattact ctagtttcgt taatatggag agagatctag cttcaggact cattggccct
     2401 ctcctcatct gctacaaaga atctgtagat caaagaggaa accagataat gtcagacaag
     2461 aggaatgtca tcctgttttc tgtatttgat gagaaccgaa gctggtacct cacagagaat
     2521 atacaacgct ttctccccaa tccagctgga gtgcagcttg aggatccaga gttccaagcc
     2581 tccaacatca tgcacagcat caatggctat gtttttgata gtttgcagtt gtcagtttgt
     2641 ttgcatgagg tggcatactg gtacattcta agcattggag cacagactga cttcctttct
     2701 gtcttcttct ctggatatac cttcaaacac aaaatggtct atgaagacac actcacccta
     2761 ttcccattct caggagaaac tgtcttcatg tcgatggaaa acccaggtct atggattctg
     2821 gggtgccaca actcagactt tcggaacaga ggcatgaccg ccttactgaa ggtttctagt
     2881 tgtgacaaga acactggtga ttattacgag gacagttatg aagatatttc agcatacttg
     2941 ctgagtaaaa acaatgccat tgaaccaaga agcttctctc aaaacccacc agtcttgaaa
     3001 cgccatcaac gggaaataac tcgtactact cttcagtcag atcaagagga aattgactat
     3061 gatgatacca tatcagttga aatgaagaag gaagattttg acatttatga tgaggatgaa
     3121 aatcagagcc cccgcagctt tcaaaagaaa acacgacact attttattgc tgcagtggag
     3181 aggctctggg attatgggat gagtagctcc ccacatgttc taagaaacag ggctcagagt
     3241 ggcagtgtcc ctcagttcaa gaaagttgtt ttccaggaat ttactgatgg ctcctttact
     3301 cagcccttat accgtggaga actaaatgaa catttgggac tcctggggcc atatataaga
     3361 gcagaagttg aagataatat catggtaact ttcagaaatc aggcctctcg tccctattcc
     3421 ttctattcta gccttatttc ttatgaggaa gatcagaggc aaggagcaga acctagaaaa
     3481 aactttgtca agcctaatga aaccaaaact tacttttgga aagtgcaaca tcatatggca
     3541 cccactaaag atgagtttga ctgcaaagcc tgggcttatt tctctgatgt tgacctggaa
     3601 aaagatgtgc actcaggcct gattggaccc cttctggtct gccacactaa cacactgaac
     3661 cctgctcatg ggagacaagt gacagtacag gaatttgctc tgtttttcac catctttgat
     3721 gagaccaaaa gctggtactt cactgaaaat atggaaagaa actgcagggc tccctgcaat
     3781 atccagatgg aagatcccac ttttaaagag aattatcgct tccatgcaat caatggctac
     3841 ataatggata cactacctgg cttagtaatg gctcaggatc aaaggattcg atggtatctg
     3901 ctcagcatgg gcagcaatga aaacatccat tctattcatt tcagtggaca tgtgttcact
     3961 gtacgaaaaa aagaggagta taaaatggca ctgtacaatc tctatccagg tgtttttgag
     4021 acagtggaaa tgttaccatc caaagctgga atttggcggg tggaatgcct tattggcgag
     4081 catctacatg ctgggatgag cacacttttt ctggtgtaca gcaataagtg tcagactccc
     4141 ctgggaatgg cttctggaca cattagagat tttcagatta cagcttcagg acaatatgga
     4201 cagtgggccc caaagctggc cagacttcat tattccggat caatcaatgc ctggagcacc
     4261 aaggagccct tttcttggat caaggtggat ctgttggcac caatgattat tcacggcatc
     4321 aagacccagg gtgcccgtca gaagttctcc agcctctaca tctctcagtt tatcatcatg
     4381 tatagtcttg atgggaagaa gtggcagact tatcgaggaa attccactgg aaccttaatg
     4441 gtcttctttg gcaatgtgga ttcatctggg ataaaacaca atatttttaa ccctccaatt
     4501 attgctcgat acatccgttt gcacccaact cattatagca ttcgcagcac tcttcgcatg
     4561 gagttgatgg gctgtgattt aaatagttgc agcatgccat tgggaatgga gagtaaagca
     4621 atatcagatg cacagattac tgcttcatcc tactttacca atatgtttgc cacctggtct
     4681 ccttcaaaag ctcgacttca cctccaaggg aggagtaatg cctggagacc tcaggtgaat
     4741 aatccaaaag agtggctgca agtggacttc cagaagacaa tgaaagtcac aggagtaact
     4801 actcagggag taaaatctct gcttaccagc atgtatgtga aggagttcct catctccagc
     4861 agtcaagatg gccatcagtg gactctcttt tttcagaatg gcaaagtaaa ggtttttcag
     4921 ggaaatcaag actccttcac acctgtggtg aactctctag acccaccgtt actgactcgc
     4981 taccttcgaa ttcaccccca gagttgggtg caccagattg ccctgaggat ggaggttctg
     5041 ggctgcgagg cacaggacct ctactgagcg gccgctcgag gtcacccatt cgaacaaaaa
     5101 ctcatctcag aagaggatct gaatatgcat accggtcatc atcaccatca ccattgagtt
     5161 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5221 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5281 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5341 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     5401 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     5461 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5521 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5581 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5641 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5701 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5761 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5821 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5881 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5941 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     6001 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6061 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6121 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     6181 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     6241 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     6301 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     6361 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     6421 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     6481 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     6541 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     6601 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     6661 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     6721 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6781 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6841 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6901 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6961 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7021 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7081 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7141 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7201 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9181 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     9241 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     9301 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     9361 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
