LOCUS       sequence-391143-        5197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the biliverdin-binding serpin of Boana punctata 
            treefrog (BpBBS) in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5197)
  AUTHORS   Manoilov KY, Ghosh A, Almo SC, Verkhusha VV
  TITLE     Structural and Functional Characterization of a Biliverdin-Binding 
            Near-Infrared Fluorescent Protein From the Serpin Superfamily.
  JOURNAL   J Mol Biol. 2022 Jan 30;434(2):167359. doi: 
            10.1016/j.jmb.2021.167359. Epub 2021 Nov 16.
  PUBMED    34798132
REFERENCE   2  (bases 1 to 5197)
  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 Mol 
            Biol."; date: "2022-01-30"; volume: "434(2):167359. doi"; pages: " 
            10.1016/j.jmb.2021.167359. Epub 2021 Nov 16"
COMMENT     Sequence Label: pCMV-BpBBS
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        425..628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     579..599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      678..687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2045..2166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2082..2101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2136..2155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2173..2628)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2310..2331)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2522..2541
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2655..2759
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2761..3118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2761..2952
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2784..2804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2969..3104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3153..3947
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3153..3947
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3207..3226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3817..3836
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4135..4154)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4179..4226
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4555..5143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5044..5063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      661 ccggactcag atctgccgcc accatggcgc accacgagga gggacaccac gaccacgagg
      721 atctgaagga cgatcacgac ccattcctgc ccgaggatca caagaaggcc ctgttcgtgt
      781 accagaagcc cgccctgaac aatatcaact tcgcctttaa gatgtataga cagctggcca
      841 gggaccaccc tacagagaat atcgtgatct ccccagtgtc tatcagctcc gccctggccc
      901 tgctgagcct gggagcaaag ggacacaccc actcccagat cgtggagaga ctgggctaca
      961 acacatctga gatccctgag cagcagatcc acgagagctt ccacaagcag ctggacgtgg
     1021 tggacgataa ggacagggat ctggagttcg agcacggcaa tgccctgttt acctgcaagg
     1081 agcacaagat ccaccagaca ttcctggacg atgccaagaa gttttaccac tctgaagtga
     1141 tcccaaccga cttcaagaac acagaggagg ccaagaacca gatcaatagc tatgtggaga
     1201 agtccaccca cggcaagatc acaaatatcc tggatagcgt ggaccaggat gccatgatcg
     1261 ccctgatcaa ctttatctat ctgcgcgcca attggcagca ccctttcgac gagaagctga
     1321 ccaaggaggg cgattttcac gtggacaagg ataccacagt gaaggtgcca ttcatgcgga
     1381 gaaggggcat ctacaagatg gcctataccg acgatatcat catggtgaca atcccctaca
     1441 acggctccgt ggagatgttt ctggccatga caaagatggg caagctgtcc gagctggagc
     1501 agaacctgaa tcgcgagcgg tctctgaagt ggcgggagat catgcagtat cagctgatcg
     1561 acctgagcct gcccaagctg tctgtgagcg gcatcctgaa cctgaaggag accctgtcca
     1621 agctgggcat cgtggacgtg ttcagcaatc acgccgacct gtctggcatc acagatgagt
     1681 ctcacctgaa ggtgagcaag gccatccaca aggccatgat gtcttttgac gagcacggaa
     1741 ccgaggcagc accagcaaca gcagcagagg cagatcctct gatgctgcca cctcacttca
     1801 agtttgacta ccctttcatc tttcgcgtgc aggatctgaa aaccaagaac ccactgctgg
     1861 tgggccggat cgccaatccc cagaagtgag aattctgcag tcgacggtac cgcgggcccg
     1921 ggatcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1981 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2041 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2101 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2161 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2221 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2281 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2341 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2401 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2461 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2521 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2581 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     2761 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2941 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3001 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3061 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3121 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3181 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3241 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3301 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3361 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3421 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3481 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3541 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3601 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3661 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3721 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3781 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3841 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3901 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3961 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4021 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4081 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4141 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4201 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4261 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4321 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4381 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     4441 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4501 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4561 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4621 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4681 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4741 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4801 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4861 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4921 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4981 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5041 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5101 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5161 ctttttacgg ttcctggcct tttgctggcc ttttgct
//
