LOCUS       sequence-360570-        7372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of TH rat mAb HC gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7372)
  AUTHORS   Huang KL, Zhu H
  TITLE     Heng Zhu lab antibody-expressing plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7372)
  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            "unpublished"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: TH mAb_BCN53.1.1D10_HC_Rat IgG2a
FEATURES             Location/Qualifiers
     source          1..7372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..483
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        484..687
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     638..658
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     684..708
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      789..798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2202..2790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2255..2275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2673..2684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2812..2831)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2856..2904
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3106..3534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3403..3424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3543..3563)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3548..3877
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3728..3863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3790..3809
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3944..4738
                     /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(3998..4017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4608..4627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4914..5035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4951..4970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5005..5024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5084..5100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5084..5100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5097..5119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5108..5124
                     /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(5132..5138,5139..5156,5157..5162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5177..5198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5315..5332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5486..6074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5566..5585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6245..7036,7037..7105))
                     /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     6868..6887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7106..7210)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7285..7304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
       61 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      121 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      181 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      541 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      601 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      661 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      721 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
      781 gagaattcgc caccatggaa tgtaactgga tacttccttt tattctgtca gtaacttcag
      841 gtgtctactc acaggttcag ctccagcagt ctggggctga actggcaaga cctggggctt
      901 cagtgaagtt gtcctgcaag gcttctggct acacctttac taactactgg atgcagtggg
      961 taaaacagag gcctggacag ggtctggaat gtattggggc tatttatcct ggagatggtg
     1021 atactaggta cactcagaag ttcaagggca aggccacatt gactgcagat aaatcctcca
     1081 gcacagccta catgcaactc agcagcttgg catctgagga ctctgcggtc tattactgtg
     1141 caccgggaga ggggtttgct tactggggcc aagggactct ggtcactgtc tctgcagcag
     1201 agaccacagc accatccgtg tacccactgg cacctggaac cgccctgaag agcaactcca
     1261 tggtgacact gggctgcctg gtgaagggct atttcccaga gcccgtgacc gtgacatgga
     1321 acagcggcgc cctgagctcc ggagtgcaca cctttcctgc cgtgctgcag agcggcctgt
     1381 acaccctgac atctagcgtg accgtgcctt cctctacatg gagctcccag gcagtgacct
     1441 gcaacgtggc acacccagcc tctagcacaa aggtggacaa gaagatcgtg cctagggagt
     1501 gtaatccatg cggatgtacc ggatccgagg tgtccagcgt gttcatcttt ccccctaaga
     1561 ccaaggatgt gctgaccatc acactgaccc ccaaggtgac atgcgtggtg gtggacatca
     1621 gccagaacga tcctgaggtg cgcttctcct ggtttatcga cgatgtggag gtgcacacag
     1681 cacagaccca cgcaccagag aagcagtcta atagcaccct gcggtccgtg tctgagctgc
     1741 ccatcgtgca cagagactgg ctgaacggca agacattcaa gtgcaaagtg aatagcggcg
     1801 cctttcctgc cccaatcgag aagagcatct ccaagccaga gggaacccca aggggaccac
     1861 aggtgtacac aatggcccca cccaaggagg agatgaccca gtctcaggtg agcatcacat
     1921 gtatggtgaa gggcttctac cctccagata tctataccga gtggaagatg aacggccagc
     1981 ctcaggagaa ctataagaat acacccccta ccatggacac agatggctcc tacttcctgt
     2041 attctaagct gaacgtgaag aaggagacct ggcagcaggg caataccttc acctgcagcg
     2101 tgctgcacga gggcctgcac aatcaccaca cagagaagtc tctgagccac tcccccggca
     2161 agtgaaagct taccggttag taatgagttt gatatctcga caatcaacct ctggattaca
     2221 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2281 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2341 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2401 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2461 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2521 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2581 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     2641 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2701 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2761 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     2821 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     2881 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     2941 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3001 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     3061 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3181 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3241 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     3301 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3421 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     3481 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3541 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3601 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3661 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3721 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3781 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3841 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3901 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3961 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4021 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4081 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4141 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4201 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4261 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4321 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4381 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4441 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4501 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4561 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4621 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4681 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4741 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4801 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4861 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5041 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5101 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5161 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5341 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5401 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6001 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6061 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6241 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6301 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6361 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6421 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6481 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6541 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6601 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6661 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6721 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6781 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6841 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6901 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6961 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7021 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7081 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7141 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7201 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7261 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     7321 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tt
//
