LOCUS       sequence-360571-        7390 bp    DNA     circular SYN 15-APR-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 7390)
  AUTHORS   Huang KL, Zhu H
  TITLE     Heng Zhu lab antibody-expressing plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "unpublished"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_HC_Rat IgG2a
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     662..686
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      767..776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2198..2786
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2251..2271)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2669..2680)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2808..2827)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2852..2900
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3102..3530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3189..3208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3399..3420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3539..3559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3544..3873
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3724..3859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3786..3805
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3940..4734
                     /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(3994..4013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4604..4623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4910..5031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4947..4966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5001..5020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5080..5096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5080..5096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5093..5115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5104..5120
                     /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(5128..5134,5135..5152,5153..5158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5173..5194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5311..5328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5482..6070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5562..5581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6241..7032,7033..7101))
                     /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     6864..6883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7102..7206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7281..7300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
       61 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      721 atagaagaca ccgggaccga tccagcctcc ggactctaga gaattcgcca ccatggctgt
      781 tctggtgctg ttcctctgcc tggttgcatt tccaagctgt gtcctgtccc aggtgcagct
      841 gaaggagtca ggacctgtcc tggtggcgcc ctcacagagc ctgtccatca cttgcactgt
      901 ctctgggttt tcattaacca gctatggtgt acactgggtt cgccagcctc caggaaaggg
      961 tctggagtgg ctgggagtaa tatgggctgg tggaagcaca aattataatt cagctatcat
     1021 gtccagactg agcatcagca aagacaactc caagagccaa gttttcttaa aaatgaacag
     1081 tctgcaaact gatgacacag ccatgtacta ctgtgccaaa cataaggcag ctcgggctat
     1141 gtacgatgct atggactact ggggtcaagg aacctcagtc accgtctcct cagcagagac
     1201 cacagcacca tccgtgtacc cactggcacc tggaaccgcc ctgaagagca actccatggt
     1261 gacactgggc tgcctggtga agggctattt cccagagccc gtgaccgtga catggaacag
     1321 cggcgccctg agctccggag tgcacacctt tcctgccgtg ctgcagagcg gcctgtacac
     1381 cctgacatct agcgtgaccg tgccttcctc tacatggagc tcccaggcag tgacctgcaa
     1441 cgtggcacac ccagcctcta gcacaaaggt ggacaagaag atcgtgccta gggagtgtaa
     1501 tccatgcgga tgtaccggat ccgaggtgtc cagcgtgttc atctttcccc ctaagaccaa
     1561 ggatgtgctg accatcacac tgacccccaa ggtgacatgc gtggtggtgg acatcagcca
     1621 gaacgatcct gaggtgcgct tctcctggtt tatcgacgat gtggaggtgc acacagcaca
     1681 gacccacgca ccagagaagc agtctaatag caccctgcgg tccgtgtctg agctgcccat
     1741 cgtgcacaga gactggctga acggcaagac attcaagtgc aaagtgaata gcggcgcctt
     1801 tcctgcccca atcgagaaga gcatctccaa gccagaggga accccaaggg gaccacaggt
     1861 gtacacaatg gccccaccca aggaggagat gacccagtct caggtgagca tcacatgtat
     1921 ggtgaagggc ttctaccctc cagatatcta taccgagtgg aagatgaacg gccagcctca
     1981 ggagaactat aagaatacac cccctaccat ggacacagat ggctcctact tcctgtattc
     2041 taagctgaac gtgaagaagg agacctggca gcagggcaat accttcacct gcagcgtgct
     2101 gcacgagggc ctgcacaatc accacacaga gaagtctctg agccactccc ccggcaagtg
     2161 aaagcttacc ggttagtaat gagtttgata tctcgacaat caacctctgg attacaaaat
     2221 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2281 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2341 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2401 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2461 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2521 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2581 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     2641 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2701 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2761 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     2821 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     2881 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     2941 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3001 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3061 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3241 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3721 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3781 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3841 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3901 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3961 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4021 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4081 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4141 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4201 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4261 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4321 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4381 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4441 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4501 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4561 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4621 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4681 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4741 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4801 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4861 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4921 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4981 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5041 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5101 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5161 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5221 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5281 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5341 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5401 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5461 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5521 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5581 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5641 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5701 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5761 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5821 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5881 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5941 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6001 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6061 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6121 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6181 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6241 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6301 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6361 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6421 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6481 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6541 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6601 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6661 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6721 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6781 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6841 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6901 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6961 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7021 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7081 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7141 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7201 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7261 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7321 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7381 aggcaaggct
//
