LOCUS       sequence-380463-        5392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hec1 heavy chain variable and constant region (mouse) + exogenous 
            N-term signal peptide. For co-expression with pDL002 in HEK293 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5392)
  AUTHORS   DeLuca KF, Mick JE, Ide AH, Lima WC, Sherman L, Schaller KL, 
            Anderson SM, Zhao N, Stasevich TJ, Varma D, Nilsson J, DeLuca JG
  TITLE     Generation and diversification of recombinant monoclonal antibodies.
  JOURNAL   Elife. 2021 Dec 31;10. pii: 72093. doi: 10.7554/eLife.72093.
  PUBMED    34970967
REFERENCE   2  (bases 1 to 5392)
  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: "Elife."; 
            date: "2021-12-31"; volume: "10. pii: 72093. doi"; pages: " 
            10.7554/eLife.72093"
COMMENT     Sequence Label: pDL001_Hec1-ms_IgG_HC
FEATURES             Location/Qualifiers
     source          1..5392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1339..2031
                     /codon_start=1
                     /product="CH2 and CH3 domains of the mouse IgG2a heavy 
                     chain plus the hinge region"
                     /label=Fc tag
                     /translation="PRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPMVT
                     CVVVDVSEDDPDVQISWFVNNVEVLTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEF
                     KCKVNNKALPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIY
                     VEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHT
                     TKSFSRTPG"
     CDS             2038..2055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2180..2301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2217..2236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2271..2290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2308..2763)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2445..2466)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2657..2676
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2790..2894
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2896..3253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2896..3087
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2919..2939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3104..3239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3288..4082
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3288..4082
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3342..3361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3952..3971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4270..4289)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4314..4361
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4690..5278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5179..5198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct cgccaccatg gaatggagct gggtctttct cttcttcctg
      661 tcagtaacta caggtgtcca ctccgacgtg cagctccaag agtctggccc tggcctggtt
      721 aagcctagcc agagcctgag cctgacatgt accgtgaccg gctacagcat caccagcgac
      781 tacagctgga actggatcag acagttcccc ggcaacaaac tggaatggat gggctacatc
      841 agctacagcg gcaccaccaa ctacaacccc agcctgaaaa cccggatctc catcaccaga
      901 gacaccagca agaaccagtt cttcctgcgg ctgaacagcg tgaccgccga agatacaggc
      961 acctactact gcgccatcta ctacggcaga ctgaagccct acatcgacta ctggggccag
     1021 ggaacaacac tggccgtgtc tagcgccaag acaacagccc cttctgtgta ccctctggct
     1081 cctgtgtgtg gcgatacaac aggcagctct gtgaccctgg gctgtctggt caagggctac
     1141 tttcccgagc ctgtgacact gacctggaac agcggatctc tgtctagcgg cgtgcacacc
     1201 tttccagccg tgctgcagag cgatctgtac accctgtcca gcagcgtgac agtgaccagc
     1261 agcacatggc ccagccagag catcacctgt aacgtggccc atcctgccag ctccaccaag
     1321 gtggacaaga agatcgagcc cagagggccc acaatcaagc cctgtcctcc atgcaaatgc
     1381 ccagcaccta acctcttggg tggaccatcc gtcttcatct tccctccaaa gatcaaggat
     1441 gtactcatga tctccctgag ccccatggtc acatgtgtgg tggtggatgt gagcgaggat
     1501 gacccagatg tccagatcag ctggttcgtg aacaacgtgg aagtactcac agctcagaca
     1561 caaacccata gagaggatta caacagtact ctccgggtgg tcagtgccct ccccatccag
     1621 caccaggact ggatgagtgg caaggagttc aaatgcaagg tcaacaacaa agccctccca
     1681 gcgcccatcg agagaaccat ctcaaaaccc aaagggtcag taagagctcc acaggtatat
     1741 gtcttgcctc caccagaaga agagatgact aagaaacagg tcactctgac ctgcatggtc
     1801 acagacttca tgcctgaaga catttacgtg gagtggacca acaacgggaa aacagagcta
     1861 aactacaaga acactgaacc agtcctggac tctgatggtt cttacttcat gtacagcaag
     1921 ctgagagtgg aaaagaagaa ctgggtggaa agaaatagct actcctgctc agtggtccac
     1981 gagggtctgc acaatcacca cacgactaag agcttctccc ggactccggg tctcgagcat
     2041 caccaccatc accattgagc ggccgcgact ctagatcata atcagccata ccacatttgt
     2101 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2161 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2221 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2281 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2341 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2401 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2461 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2521 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2581 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2641 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2701 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2881 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3061 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3181 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3241 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3601 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3781 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4081 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4261 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4321 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4381 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4441 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4501 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5341 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
