LOCUS       sequence-360643-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PVALB rat mAb LC gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  AUTHORS   Huang KL, Zhu H
  TITLE     Heng Zhu lab antibody-expressing plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6697)
  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            "unpublished"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_BCN14.1.1A2_LC_Rat Ig Kappa
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        436..639
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     590..610
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     636..660
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    1479..2067
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1532..1552)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1950..1961)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2089..2108)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2133..2181
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2383..2811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2820..2840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2825..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3067..3086
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3221..4015
                     /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(3275..3294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3885..3904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4191..4312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4282..4301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4361..4377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4361..4377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4374..4396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4385..4401
                     /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(4409..4415,4416..4433,4434..4439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4454..4475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4592..4609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4763..5351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4843..4862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5522..6313,6314..6382))
                     /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     6145..6164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6383..6487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6562..6581)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
       61 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      121 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      181 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      241 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      301 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      361 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      421 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      481 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      541 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      601 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
      661 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      721 ctccggactc tagagaattc gccaccatgc tcactcagct cctgggatta ctgctgctct
      781 ggtttgcagg tggtaaatgt gacattcaga tgacccagtc tcctgcctcc cagtctgcat
      841 ctctgggaga aagtgtcacc atcacatgcc tggcaagtca gaccattggt acatggttag
      901 catggtatca gcagaaacca gggaaatctc ctcagctcct gatttatgct gcaaccagct
      961 tggcagatgg ggtcccatca aggttcagtg gtagtggatc tggcacaaag ttttctttca
     1021 agatcagcag cctacaggct gaagattttg caagttatta ctgtcaacaa cattacatta
     1081 ctcctctgac gttcggtgga ggcaccaagc tggaaatcaa agccgacgcc gccccaaccg
     1141 tgtctatctt cccccctagc atggagcagc tgacctctgg cggcgccaca gtggtgtgct
     1201 tcgtgaacaa cttctacccc cgggacatct ccgtgaagtg gaagatcgat ggctctgagc
     1261 agagagacgg cgtgctggat agcgtgaccg accaggattc taaggatagc acatattcca
     1321 tgagctccac cctgtccctg acaaaggtgg agtacgagag gcacaacctg tatacctgcg
     1381 aggtggtgca caagacatct agctcccccg tggtgaagtc ctttaaccgg aatgagtgtt
     1441 gaaagcttac cggttagtaa tgagtttgat atctcgacaa tcaacctctg gattacaaaa
     1501 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     1561 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     1621 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     1681 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     1741 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     1801 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1861 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     1921 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     1981 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     2041 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     2101 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     2161 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     2221 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     2281 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     2341 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     2761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2821 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3181 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3241 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3301 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3361 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3421 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3481 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3541 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3601 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3661 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3721 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3781 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3841 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3901 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3961 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4021 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4081 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4201 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4261 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4321 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4501 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4561 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4621 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4681 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4741 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4801 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5101 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5161 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5221 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5281 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5341 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5401 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5461 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5521 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5641 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5821 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6361 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6481 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6541 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6601 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6661 aaggcaaggc ttgaccgaca attgcatgaa gaatctg
//
