LOCUS       sequence-360565-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PVALB rat mAb HC gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Huang KL, Zhu H
  TITLE     BICAN Project Antibody Sequences
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7369)
  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            "unpublished"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: PVALB mAb_BCN14.1.1C9_HC_Rat IgG2a
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     631..655
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      736..745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2146..2734
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2199..2219)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2617..2628)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2756..2775)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2800..2848
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3050..3478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3347..3368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3487..3507)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3492..3821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3672..3807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3734..3753
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3888..4682
                     /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(3942..3961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4552..4571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4858..4979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4895..4914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4949..4968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5028..5044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5028..5044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5041..5063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5052..5068
                     /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(5076..5082,5083..5100,5101..5106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5121..5142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5259..5276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5430..6018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5510..5529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6189..6980,6981..7049))
                     /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     6812..6831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7050..7154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7229..7248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
       61 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      661 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      721 gactctagag aattcgccac catggactcc aggctcaatt tagttttcct tgtccttatt
      781 ttaaaaggtg tccagtgtga ggtgcagctg gtggagtctg ggggaggctt agtgaagcct
      841 ggagggtccc ggaaactctc ctgtgcagcc tctggattca ctttcagtga ctatggaatg
      901 ctctgggtcc gtcaggctcc agagaagggg ctggagtggg ttgcatacat tagtagtggc
      961 agtagtacca tctactatgc agacacagtg aagggccgat tcaccatctc cagagacaat
     1021 gccaagaaca ccctgttcct gcaaatgacc agtctaaggt ctgaggacac agccgtgtat
     1081 tactgtgcaa gggctgggac cgactactgg ggccaaggca ccactctcac agtctcctca
     1141 gcagagacca cagcaccatc cgtgtaccca ctggcacctg gaaccgccct gaagagcaac
     1201 tccatggtga cactgggctg cctggtgaag ggctatttcc cagagcccgt gaccgtgaca
     1261 tggaacagcg gcgccctgag ctccggagtg cacacctttc ctgccgtgct gcagagcggc
     1321 ctgtacaccc tgacatctag cgtgaccgtg ccttcctcta catggagctc ccaggcagtg
     1381 acctgcaacg tggcacaccc agcctctagc acaaaggtgg acaagaagat cgtgcctagg
     1441 gagtgtaatc catgcggatg taccggatcc gaggtgtcca gcgtgttcat ctttccccct
     1501 aagaccaagg atgtgctgac catcacactg acccccaagg tgacatgcgt ggtggtggac
     1561 atcagccaga acgatcctga ggtgcgcttc tcctggttta tcgacgatgt ggaggtgcac
     1621 acagcacaga cccacgcacc agagaagcag tctaatagca ccctgcggtc cgtgtctgag
     1681 ctgcccatcg tgcacagaga ctggctgaac ggcaagacat tcaagtgcaa agtgaatagc
     1741 ggcgcctttc ctgccccaat cgagaagagc atctccaagc cagagggaac cccaagggga
     1801 ccacaggtgt acacaatggc cccacccaag gaggagatga cccagtctca ggtgagcatc
     1861 acatgtatgg tgaagggctt ctaccctcca gatatctata ccgagtggaa gatgaacggc
     1921 cagcctcagg agaactataa gaatacaccc cctaccatgg acacagatgg ctcctacttc
     1981 ctgtattcta agctgaacgt gaagaaggag acctggcagc agggcaatac cttcacctgc
     2041 agcgtgctgc acgagggcct gcacaatcac cacacagaga agtctctgag ccactccccc
     2101 ggcaagtgaa agcttaccgg ttagtaatga gtttgatatc tcgacaatca acctctggat
     2161 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2221 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2281 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2341 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2401 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2461 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2521 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2581 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2641 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2701 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     2761 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2821 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2881 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2941 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     3001 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3421 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3481 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3541 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3661 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3721 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3781 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3841 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4741 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4801 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4861 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4921 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4981 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5041 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5101 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5161 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5221 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5281 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5341 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5401 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5461 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5521 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5581 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5641 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5701 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5761 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5821 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5881 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5941 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6001 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6061 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6121 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6181 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6241 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6301 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6361 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6421 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6481 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6541 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6601 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6661 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6721 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6781 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6841 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6901 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6961 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7021 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7081 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7141 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7201 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     7261 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7321 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttag
//
