LOCUS       sequence-377349-        8003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8003)
  AUTHORS   Yen TJ, Lolicato M, Thomas-Tran R, Du Bois J, Minor DL Jr
  TITLE     Structure of the saxiphilin:saxitoxin (STX) complex reveals a 
            convergent molecular recognition strategy for paralytic toxins.
  JOURNAL   Sci Adv. 2019 Jun 19;5(6):eaax2650. doi: 10.1126/sciadv.aax2650. 
            eCollection 2019 Jun.
  PUBMED    31223657
REFERENCE   2  (bases 1 to 8003)
  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: "Sci Adv."; 
            date: "2019-06-19"; volume: "5(6):eaax2650. doi"; pages: " 
            10.1126/sciadv.aax2650. eCollection 2019 Jun"
COMMENT     Sequence Label: pFastBac1-bullfrog saxiphilin
FEATURES             Location/Qualifiers
     source          1..8003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        239..330
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     255..272
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2917..2940
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2956..2958,2959..3669)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2998..3019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3259..3278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3606..3627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3682..3711
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    3847..3981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3884..3903)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3938..3957
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4010..4175
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4359..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4446..4465)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4655..4676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4839..4943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4944..5012,5013..5804)
                     /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     complement(5162..5181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5975..6563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6464..6483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6717..6734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6868..7092
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7159..7692)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7862..7867,7868..7884,7885..7890))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg
       61 atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc
      121 tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg
      181 cccaggactc tagctatagt tctagtggtt ggctacgtat actccggaat attaatagat
      241 catggagata attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt
      301 aacagttttg taataaaaaa acctataaat attccggatt attcataccg tcccaccatc
      361 gggcgcggat ccatggctcc taccttccag accgctctgt tcttcaccat catctccctg
      421 tccttcgcta tggccccgaa cgcaaagcaa gtccgctggt gtgctatcag cgacctggag
      481 cagaagaaat gcaacgatct ggttggtagc tgcaacgtgc cggacatcac cctggtgtgc
      541 gttctgcgta gcagcaccga agattgcatg accgcgatca aggacggtca ggcggatgct
      601 atgttcctgg acagcggcga ggtgtacgaa gcgagcaaag atccgtacaa cctgaagccg
      661 atcatcgctg aaccgtacag cagcaaccgt gacctgcaga agtgcctgaa agagcgtcag
      721 caggcgctgg ctaagaaaat gatcggtcac tacatcccgc agtgcgatga gaaaggcaac
      781 taccaaccgc agcagtgcca cggtagcacc ggtcactgct ggtgcgttaa cgcgatgggt
      841 gaaaagatca gcggtaccaa caccccgccg ggtcagaccc gtgctacctg cgagcgtcac
      901 gaactgccga agtgcctgaa agaacgtcag gtggcgctgg gtggcgacga gaaagttctg
      961 ggtcgtttcg tgccgcagtg cgatgaaaag ggcaactacg agccgcagca gttccacggt
     1021 agcaccggct actcctggtg tgttaacgcg atcggcgagg aaatcgctgg caccaaaacc
     1081 ccgccgggca agatcccggc tacctgccag aagcacgacc tggtgaccac ctgccactac
     1141 gcggttgcta tggtgaagaa aagcagcgcg ttccagttca accagctgaa gggcaaacgt
     1201 agctgccaca gcggtgttag caaaaccgac ggctggaagg cgctggtgac cgttctggtg
     1261 gaaaagaaac tgctgagctg ggatggtccg gcgaaagaga gcatccagcg tgctatgagc
     1321 aagttcttca gcgtgagctg cattccgggt gctacccaga ccaacctgtg caagcagtgc
     1381 aaaggcgagg aaggcaagaa ctgcaaaaac agccacgacg aaccgtacta cggtaactac
     1441 ggcgcgttcc gttgcctgaa ggaagacatg ggtgacgttg cgttcctgcg tagcaccgct
     1501 ctgagcgacg agcacagcga agtgtacgag ctgctgtgcc cggataacac ccgtaagccg
     1561 ctgaacaagt acaaagagtg caacctgggt accgttccgg ctggcaccgt ggttacccgt
     1621 aagatcagcg acaaaaccga agacatcaac aacttcctga tggaggcgca gaagcgtcag
     1681 tgcaaactgt tcagcagcgc tcacggcaag gacctgatgt tcgacgatag caccctgcag
     1741 ctggctctgc tgagcagcga ggttgatgct ttcctgtacc tgggtgtgaa actgttccac
     1801 gcgatgaagg ctctgaccgg cgacgctcac ctgccgagca agaacaaagt gcgttggtgc
     1861 accatcaaca agctggaaaa gatgaaatgc gacgattgga gcgcggttag cggtggcgcg
     1921 atcgcttgca ccgaggctag ctgcccgaag ggctgcgtga aacagatcct gaagggcgaa
     1981 gcggacgctg ttaagctgga agtgcagtac atgtacgagg cgctgatgtg cggtctgctg
     2041 ccggctgttg aggaatacca caacaaagac gatttcggtc cgtgcaagac cccaggtagc
     2101 ccgtacaccg acttcggcac cctgcgtgcg gttgctctgg tgaagaaaag caacaaagac
     2161 atcaactgga acaacatcaa gggcaagaag agctgccaca ccggtgtggg cgacatcgct
     2221 ggctgggtta tcccggtgag cctgatccgt cgtcagaacg ataacagcga catcgatagc
     2281 ttcttcggcg agagctgcgc gccgggcagc gacaccaaaa gcaacctgtg caagctgtgc
     2341 atcggtgacc cgaagaacag cgcggctaac accaaatgca gcctgagcga caaggaagcg
     2401 tactacggta accagggcgc tttccgttgc ctggtggaga aaggtgacgt tgctttcgtg
     2461 ccgcacaccg tggttttcga aaacaccgat ggcaagaacc cggcggtttg ggctaagaac
     2521 ctgaaaagcg aagacttcga gctgctgtgc ctggatggta gccgtgcgcc ggtgagcaac
     2581 tacaagagct gcaaactgag cggtatcccg ccgccggcta tcgttacccg tgaggaaagc
     2641 atcagcgacg tggttcgtat cgtggctaac cagcagagcc tgtacggtcg taaaggcttc
     2701 gagaaggata tgttccagct gttcagcagc aacaagggta acaacctgct gttcaacgac
     2761 aacacccagt gcctgatcac cttcgatcgt cagccgaaag acatcatgga agattacttc
     2821 ggcaagccgt actacaccac cgtgtacggc gcgtcccgct ccgctatgtc ctccgaactg
     2881 atttccgcct gtaccattaa acactgctcg aattctttgg aagttttgtt tcaaggtcca
     2941 actgctgccg ccgctgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3001 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3061 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3121 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3181 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3241 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3301 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3361 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3421 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3481 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3541 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     3601 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3661 ctgtacaagt ccggaggtgg tcaccatcac catcaccacc atcaccatca ctaatagaag
     3721 cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga ggttttactt
     3781 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3841 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3901 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3961 gtatcttatc atgtctggat ctgatcactg cttgagccta ggagatccga accagataag
     4021 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     4081 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     4141 cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg
     4201 tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc
     4261 tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg
     4321 aatatcaacg cttatttgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     4381 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4441 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4501 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4561 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4621 tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca aactggaaca
     4681 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4741 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4801 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4921 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4981 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5041 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5101 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5161 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5221 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5281 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5341 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5401 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5461 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5521 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5581 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5641 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5701 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5761 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5821 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5881 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6661 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6721 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     6781 gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt acggttgagt
     6841 aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta aactgaacaa
     6901 aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat
     6961 cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt
     7021 cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa gggcatggta
     7081 aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat
     7141 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
     7201 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
     7261 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
     7321 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
     7381 actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
     7441 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
     7501 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
     7561 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
     7621 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     7681 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     7741 catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     7801 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     7861 cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg
     7921 tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt cctggctggc
     7981 gaacgagcgc aaggtttcgg tct
//
