LOCUS       sequence-156201-        5733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chain A of the habu snake venom FIX/FX binding protein with a 
            C-terminal polyhistidine-tag. Cloned into pIRESneo3 (Ampicillin) via
            5' NheI and 3' BamHI; IRES vector conferring neomycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5733)
  AUTHORS   Robert Peters
  TITLE     rFIX/X-binding protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5733)
  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            "Unpublished"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: rFIXX-bp chain A (IRESneo)
FEATURES             Location/Qualifiers
     source          1..5733
                     /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      934..943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1408..1431
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     intron          1509..1738
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1721..1740
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1811..2385,2386..2388,2389..2397)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1978..1995)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2205..2224
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2420..3223
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2483..2502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3093..3112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3353..3474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3390..3409)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3576..3592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3576..3592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3589..3611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3600..3616
                     /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(3624..3630,3631..3648,3649..3654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3669..3690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3807..3824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3978..4566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4058..4077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4737..5528,5529..5597))
                     /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     5360..5379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5598..5702)
                     /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 gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gccgccacca tggggcgatt catcttcatg
      961 agcttcggct tgctggtcgt ggctgcctcc ctaagaggaa ctggagctga ttgtctctct
     1021 ggttggtcct cctatgaagg gcattgctac aaggccttcg aaaaatacaa gacctgggaa
     1081 gatgcagaga gggtctgcac ggagcaggcg aagggcgcgc atctggtctc tatcgaaagc
     1141 tccggagaag cagactttgt ggcccagctg gtcactcaga acatgaagag actcgatttc
     1201 tatatctgga tcgggctgag ggttcaaggc aaagtcaagc aatgcaactc ggagtggagc
     1261 gatggctcca gcgtcagtta tgagaactgg attgaagcag aatccaaaac gtgtcttggg
     1321 ctggaaaaag agacagattt tcgtaagtgg gtcaatattt actgtggaca acaaaatcct
     1381 ttcgtctgcg aggcaggagg aggcagccac catcaccacc atcatcacca ttgaggatcc
     1441 gcggccgcat agataactga tccagtgtgc tggaattaat tcgctgtctg cgagggccag
     1501 ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa gattgtcagt
     1561 ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt tgagggtggc
     1621 cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt gtggcaggct
     1681 tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct ctccacaggt
     1741 gtccactccc aggtccaact gcaggtcgag catgcatcta gggcggccaa ttccgcccct
     1801 ctcccccccc cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     1861 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     1921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1981 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2341 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccgg
     2401 gataattcct gcagccaata tgggatcggc cattgaacaa gatggattgc acgcaggttc
     2461 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2521 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2581 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2641 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2701 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2761 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2821 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2881 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2941 cgccaggctc aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc
     3001 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3061 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3121 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3181 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaggggatc aattctctag
     3241 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3301 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3361 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3421 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3481 cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     3541 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3601 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3661 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3721 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3781 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4381 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4621 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4681 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4741 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4801 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4861 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4921 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4981 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5041 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5101 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5161 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5221 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5281 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5341 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5401 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5461 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5521 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5581 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5641 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5701 cgcacatttc cccgaaaagt gccacctgac gtc
//
