LOCUS       sequence-57346-e        8280 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 8280)
  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 8280)
  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: pSYN-PC5-003
FEATURES             Location/Qualifiers
     source          1..8280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      913..922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3705..3722)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3711..3935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3981..4409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4087)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4278..4299
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4418..4438)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4423..4752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4603..4738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4665..4684
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4801..5826
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4801..5826
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5956..6077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5993..6012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6047..6066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6126..6142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6126..6142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6139..6161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6150..6166
                     /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(6174..6180,6181..6198,6199..6204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6219..6240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6357..6374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6528..7113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6608..6627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7284..8075,8076..8144))
                     /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     7907..7926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8145..8249)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat gggctggggg agccgctgct gctgcccggg acgtttggac
      961 ctgctgtgcg tgctggcgct gctcgggggc tgcctgctcc ccgtgtgtcg gacgcgcgtc
     1021 tacaccaacc actgggcagt caaaatcgcc gggggcttcc cggaggccaa ccgtatcgcc
     1081 agcaagtacg gattcatcaa cataggacag ataggggccc tgaaggacta ctaccacttc
     1141 taccatagca ggacgattaa aaggtcagtt atctcgagca gagggaccca cagtttcatt
     1201 tcaatggaac caaaggtgga atggatccaa cagcaagtgg taaaaaagcg gacaaagagg
     1261 gattatgact tcagtcgtgc ccagtctacc tatttcaatg atcccaagtg gcccagtatg
     1321 tggtatatgc actgcagtga caatacacat ccctgccagt ctgacatgaa tatcgaagga
     1381 gcctggaaga gaggctacac gggaaagaac attgtggtca ctatcctgga tgacggaatt
     1441 gagagaaccc atccagatct gatgcaaaac tacgatgctc tggcaagttg cgacgtgaat
     1501 gggaatgact tggacccaat gcctcgttat gatgcaagca acgagaacaa gcatgggact
     1561 cgctgtgctg gagaagtggc agccgctgca aacaattcgc actgcacagt cggaattgct
     1621 ttcaacgcca agatcggagg agtgcgaatg ctggacggag atgtcacgga catggttgaa
     1681 gcaaaatcag ttagcttcaa cccccagcac gtgcacattt acagcgccag ctggggcccg
     1741 gatgatgatg gcaagactgt ggacggacca gcccccctca cccggcaagc ctttgaaaac
     1801 ggcgttagaa tggggcggag aggcctcggc tctgtgtttg tttgggcatc tggaaatggt
     1861 ggaaggagca aagaccactg ctcctgtgat ggctacacca acagcatcta caccatctcc
     1921 atcagcagca ctgcagaaag cggaaagaaa ccttggtacc tggaagagtg ttcatccacg
     1981 ctggccacaa cctacagcag cggggagtcc tacgataaga aaatcatcac tacagatctg
     2041 aggcagcgtt gcacggacaa ccacactggg acgtcagcct cagcccccat ggctgcaggc
     2101 atcattgcgc tggccctgga agccaatccg tttctgacct ggagagacgt acagcatgtt
     2161 attgtcagga cttcccgtgc gggacatttg aacgctaatg actggaaaac caatgctgct
     2221 ggttttaagg tgagccatct ttatggattt ggactgatgg acgcagaagc catggtgatg
     2281 gaggcagaga agtggaccac cgttccccgg cagcacgtgt gtgtggagag cacagaccga
     2341 caaatcaaga caatccgccc taacagtgca gtgcgctcca tctacaaagc ctcaggctgc
     2401 tcagataacc ccaaccgcca tgtcaactac ctggagcacg tcgttgtgcg catcaccatc
     2461 acccacccca ggagaggaga cctggccatc tacctgacct cgccctctgg aactaggtct
     2521 cagcttttgg ccaacaggct atttgatcac tccatggaag gattcaaaaa ctgggagttc
     2581 atgaccattc attgctgggg agaaagagct gctggtgact gggtccttga agtttatgat
     2641 actccctctc agctaaggaa ctttaagact ccaggtaaat tgaaagaatg gtctttggtc
     2701 ctctacggca cctccgtgca gccatattca ccaaccaatg aatttccgaa agtggaacgg
     2761 ttccgctata gccgagttga agaccccaca gacgactatg gcacagagga ttatgcaggt
     2821 ccctgcgacc ctgagtgcag tgaggttggc tgtgacgggc caggaccaga ccactgcaat
     2881 gactgtttgc actactacta caagctgaaa aacaatacca ggatctgtgt ctccagctgc
     2941 ccccctggcc actaccacgc cgacaagaag cgctgcagga agtgtgcccc caactgtgag
     3001 tcctgctttg ggagccatgg tgaccaatgc atgtcctgca aatatggata ctttctgaat
     3061 gaagaaacca acagctgtgt tactcactgc cctgatgggt catatcagga taccaagaaa
     3121 aatctttgcc ggaaatgcag tgaaaactgc aagacatgta ctgaattcca taactgtaca
     3181 gaatgtaggg atgggttaag cctgcaggga tcccggtgct ctgtctcctg tgaagatgga
     3241 cggtatttca acggccagga ctgccagccc tgccaccgct tctgcgccac ttgtgctggg
     3301 gcaggagctg atgggtgcat taactgcaca gagggctact tcatggagga tgggagatgc
     3361 gtgcagagct gtagtatcag ctattacttt gaccactctt cagagaatgg atacaaatcc
     3421 tgcaaaaaat gtgatatcag ttgtttgacg tgcaatggcc caggattcaa gaactgtaca
     3481 agctgcccta gtgggtatct cttagactta ggaatgtgtc aaatgggagc catttgcaag
     3541 gatgcaacgg aagagtcctg ggcggaagga ggcttctgta tgcttgtgaa aaagaacaat
     3601 ctgtgccaac ggaaggttct tcaacaactt tgctgcaaaa catgtacatt tcaaggctga
     3661 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3721 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3781 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3841 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3901 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3961 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4021 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4081 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4141 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4201 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4261 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4321 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4381 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4501 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4561 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4621 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4681 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4741 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     4801 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     4861 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     4921 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     4981 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     5041 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     5101 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     5161 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     5221 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     5281 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     5341 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     5401 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     5461 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     5521 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     5581 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     5641 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     5701 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     5761 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     5821 gaatagcacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5881 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5941 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6061 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     7081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7561 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
