LOCUS       sequence-388075-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing an antigen targeting the HCN3 ion channel 
            subunit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  TITLE     Trimmer lab antigen plasmids
REFERENCE   2  (bases 1 to 7979)
  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"
COMMENT     Sequence Label: HCN3/pCDNA3.1
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..95)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        80..409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      260..395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     322..341
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             476..1270
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(530..549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1140..1159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1444..1565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1481..1500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1535..1554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1614..1630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1614..1630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1627..1649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1638..1654
                     /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(1662..1668,1669..1686,1687..1692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1707..1728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1845..1862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2016..2604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2096..2115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2775..3566,3567..3635))
                     /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     3398..3417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3636..3740)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3815..3834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4006..4385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4386..4589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4540..4560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4634..4653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4634..4652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4669..4678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7303..7321)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7304..7321)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7341..7358)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7347..7571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7617..7979,1..66)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7704..7723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7914..7935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
       61 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
      121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      421 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
      481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     1021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     1081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     1141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1261 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     1321 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1381 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1441 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1561 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     1621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagttcacgc
     2221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2761 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2821 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2881 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2941 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3001 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3061 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3121 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3181 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3241 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3301 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3361 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3421 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3481 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3541 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3601 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3661 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3721 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     3781 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3841 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     3901 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     3961 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     4021 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4081 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4141 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4201 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4261 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4321 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4381 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4441 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4501 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4561 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     4621 ggcttatcga aattaatacg actcactata gggagaccca agcttgccgc caccatggag
     4681 gaggaggcgc ggccggcggc gggggccggc gaagcggcga cccctgcacg cgagacgcct
     4741 cctgcggctc cggcccaggc ccgcgcggcc tcaggtgggg tgccggagtc tgcgcccgag
     4801 ccgaagaggc ggcagctcgg gacgctgctg cagccgacgg tcaacaagtt ctctctccgg
     4861 gtcttcggca gccacaaagc agtagaaatc gagcaggaga gggtgaagtc cgccggggcc
     4921 tggatcatcc acccctacag cgacttccgg ttttactggg atctcatcat gctgctgctg
     4981 atggtgggga acctcatagt tctgcctgtg ggtatcactt tcttcaagga ggagaactct
     5041 ccaccctgga tcgtcttcaa tgtcctctct gacactttct tcctgctgga tctggtgctc
     5101 aacttccgaa ctggcatcgt ggtggaggaa ggtgccgaga tcctgctggc gccaagggcc
     5161 atccgaacgc gttacctgcg cacctggttc ctggttgatc tgatctcctc catccctgtg
     5221 gattatatct tcctagtggt ggagctggag ccacgactag atgctgaggt ctacaaaacg
     5281 gcacgggccc tgcgcatcgt tagattcacc aagatcctta gcctgctgcg gctgctccgc
     5341 ctctcccgcc tcatccgcta catacaccag tgggaggaga tctttcacat gacctacgac
     5401 ctggccagtg cagtggttcg catcttcaac ctcattggaa tgatgttgct gctgtgtcac
     5461 tgggacggct gtctgcagtt tctggtccct atgctgcagg acttcccgtc cgactgctgg
     5521 gtctccatga accgcatggt gaaccactcg tggggccgcc agtattccca cgccctgttc
     5581 aaggccatga gtcacatgct atgcattggc tatgggcagc aggcaccggt aggcatgcct
     5641 gacgtctggc tcaccatgct cagtatgatt gtgggcgcca cgtgttatgc catgttcatc
     5701 ggtcacgcca ccgccctcat ccagtccctg gactcttccc ggcgacagta ccaggagaag
     5761 tacaagcagg tggagcagta catgtccttc cacaagctgc ccgctgacac ccggcagcgc
     5821 atccacgagt actacgagca tcgctaccag ggcaagatgt ttgatgaaga gagcatcctg
     5881 ggggagctga gcgagccact tcgggaggag attattaact tcacctgccg gggcctggtg
     5941 gcccacatgc cgctgtttgc tcatgctgac cccagcttcg tcaccgcagt gctcaccaag
     6001 ctccgttttg aggtcttcca accaggggac ctggtggtgc gtgagggctc cgtgggcagg
     6061 aagatgtact tcatccagca cgggctgctg agtgtgctgg cacgtggcgc ccgcgacacc
     6121 cgcctcactg atggatccta ctttggggag atctgcctgc tgactcgagg tcggagaaca
     6181 gccagtgtaa gggctgacac ctattgtcgc ctctactcgc tcagcgtgga ccacttcaat
     6241 gcggtgcttg aggagttccc aatgatgcgc agggcttttg agacggtggc catggaccgg
     6301 cttcggcgca tcggcaaaaa gaattcgata ctgcagcgga aacgctctga gccgagtcca
     6361 ggcagcagcg gtggcgtcat ggagcagcat ttggtacaac acgacagaga catggctcgt
     6421 ggtgttcggg gcctggctcc tggtacagga gctcgactca gtggaaagcc agtgctgtgg
     6481 gaaccactgg tgcacgcccc tctgcaggca gctgctgtga cctccaacgt ggccatagcc
     6541 ttgactcacc agcgaggccc tctgcccctc tcccctgatt ctccagccac cctcctagct
     6601 cgatctgcta gacgctcagc aggctcccca gcctccccac tggtgcctgt ccgagcaggt
     6661 cctctgctgg cccggggacc ctgggcgtcc acttctcgcc tgcctgctcc acctgcccga
     6721 accctccatg ccagcctatc ccggacaggg cgttcccagg tatctctgtt gggccctccc
     6781 ccaggaggag gtgctcggag gctaggacct cggggccgcc cactttctgc ctcgcaaccc
     6841 tctctgcctc agcgagcaac aggggatggc tctcctaggc gtaaaggctc tggaagtgag
     6901 cgcctgcccc cctctgggct cttggccaaa cctccaggga cagtccagcc acccaggtca
     6961 tcagtgcctg agccagttac ccccagaggt ccccaaattt ctgccaacat gtgaaatcct
     7021 aggttacact ggccttagct attggcacgc aataacgtgt gcccgagggt agacgcccct
     7081 tgggggaagg ctgtggggga cctaacatac tgccctttat caccacaaag ctattctctg
     7141 tgtcctcagc tcaaagcagc tgcagcctgc taatcctaat cacataccca tttgtcattg
     7201 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattctg cagatatcca
     7261 tcacactggc ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa
     7321 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     7381 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     7441 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     7501 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     7561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     7621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     7681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     7741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     7801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     7861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     7921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataag
//
