LOCUS       sequence-57352-e        7261 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 7261)
  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 7261)
  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-PACE-001
FEATURES             Location/Qualifiers
     source          1..7261
                     /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      2217..2226
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3125..3166
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3176..3193
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3216..3233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3222..3446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3492..3920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3579..3598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3789..3810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3929..3949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3934..4264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4115..4250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4177..4196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4312..4359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4378..4776
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4934..5055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4971..4990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5025..5044
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5104..5120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5104..5120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5117..5139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5128..5144
                     /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(5152..5158,5159..5176,5177..5182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5197..5218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5335..5352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5506..6094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5586..5605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6265..7056,7057..7125))
                     /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     6888..6907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7126..7230)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 aagcttgcca tggagctgag gccctggttg ctatgggtgg tagcagcaac
      961 aggaaccttg gtcctgctag cagctgatgc tcagggccag aaggtcttca ccaacacgtg
     1021 ggctgtgcgc atccctggag gcccagcggt ggccaacagt gtggcacgga agcatgggtt
     1081 cctcaacctg ggccagatct tcggggacta ttaccacttc tggcatcgag gagtgacgaa
     1141 gcggtccctg tcgcctcacc gcccgcggca cagccggctg cagagggagc ctcaagtaca
     1201 gtggctggaa cagcaggtgg caaagcgacg gactaaacgg gacgtgtacc aggagcccac
     1261 agaccccaag tttcctcagc agtggtacct gtctggtgtc actcagcggg acctgaatgt
     1321 gaaggcggcc tgggcgcagg gctacacagg gcacggcatt gtggtctcca ttctggacga
     1381 tggcatcgag aagaaccacc cggacttggc aggcaattat gatcctgggg ccagttttga
     1441 tgtcaatgac caggaccctg acccccagcc tcggtacaca cagatgaatg acaacaggca
     1501 cggcacacgg tgtgcggggg aagtggctgc ggtggccaac aacggtgtct gtggtgtagg
     1561 tgtggcctac aacgcccgca ttggaggggt gcgcatgctg gatggcgagg tgacagatgc
     1621 agtggaggca cgctcgctgg gcctgaaccc caaccacatc cacatctaca gtgccagctg
     1681 gggccccgag gatgacggca agacagtgga tgggccagcc cgcctcgccg aggaggcctt
     1741 cttccgtggg gttagccagg gccgaggggg gctgggctcc atctttgtct gggcctcggg
     1801 gaacgggggc cgggaacatg acagctgcaa ctgcgacggc tacaccaaca gtatctacac
     1861 gctgtccatc agcagcgcca cgcagtttgg caacgtgccg tggtacagcg aggcctgctc
     1921 gtccacactg gccacgacct acagcagtgg caaccagaat gagaagcaga tcgtgacgac
     1981 tgacttgcgg cagaagtgca cggagtctca cacgggcacc tcagcctctg cccccttagc
     2041 agccggcatc attgctctca ccctggaggc caataagaac ctcacatggc gggacatgca
     2101 acacctggtg gtacagacct cgaagccagc ccacctcaat gccaacgact gggccaccaa
     2161 tggtgtgggc cggaaagtga gccactcata tggctacggg cttttggacg caggcgccat
     2221 ggtggccctg gcccagaatt ggaccacagt ggccccccag cggaagtgca tcatcgacat
     2281 cctcaccgag cccaaagaca tcgggaaacg gctcgaggtg cggaagaccg tgaccgcgtg
     2341 cctgggcgag cccaaccaca tcactcggct ggagcacgct caggcgcggc tcaccctgtc
     2401 ctataatcgc cgtggcgacc tggccatcca cctggtcagc cccatgggca cccgctccac
     2461 cctgctggca gccaggccac atgactactc cgcagatggg tttaatgact gggccttcat
     2521 gacaactcat tcctgggatg aggatccctc tggcgagtgg gtcctagaga ttgaaaacac
     2581 cagcgaagcc aacaactatg ggacgctgac caagttcacc ctcgtactct atggcaccgc
     2641 ccctgagggg ctgcccgtac ctccagaaag cagtggctgc aagaccctca cgtccagtca
     2701 ggcctgtgtg gtgtgcgagg aaggcttctc cctgcaccag aagagctgtg tccagcactg
     2761 ccctccaggc ttcgcccccc aagtcctcga tacgcactat agcaccgaga atgacgtgga
     2821 gaccatccgg gccagcgtct gcgccccctg ccacgcctca tgtgccacat gccaggggcc
     2881 ggccctgaca gactgcctca gctgccccag ccacgcctcc ttggaccctg tggagcagac
     2941 ttgctcccgg caaagccaga gcagccgaga gtccccgcca cagcagcagc cacctcggct
     3001 gcccccggag gtggaggcgg ggcaacggct gcgggcaggg ctgctgccct cacacctgcc
     3061 tgagtgagaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggccctt
     3121 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     3181 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3841 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3961 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4021 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4081 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4141 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4201 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4261 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     4321 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     4381 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     4441 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     4501 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     4561 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     4621 gagaacaggg gcatcttgag cccctgcgga cggtgtcgac aggtgcttct cgatctgcat
     4681 cctgggatca aagcgatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     4741 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     4801 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4861 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4921 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4981 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5041 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5701 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5941 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7261 c
//
