LOCUS       sequence-380458-        4674 bp    DNA     circular SYN 14-APR-2025
DEFINITION  HA-tag light chain hypervariable regions grafted into 15F11 scaffold
            (Zhao et al., 2019) + rabbit light chain constant regions + 
            exogenous N-term signal peptide. For co-expression with pDL025..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4674)
  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 4674)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pDL026_HA-rb_IgG_LC_15F11
FEATURES             Location/Qualifiers
     source          1..4674
                     /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"
     polyA_signal    1462..1583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1499..1518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1553..1572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1590..2045)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1727..1748)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1939..1958
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2072..2176
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2178..2535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2178..2369
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2201..2221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2386..2521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2570..3364
                     /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             2570..3364
                     /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(2624..2643)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3234..3253
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3552..3571)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3596..3643
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3972..4560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4461..4480
                     /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 ggtgtgccca ctcaggtcct ggggttgctg
      661 ctgctgtggc ttacagatgc cagatgtgac attgtgctga cccaatctcc agcttctttg
      721 actgtgtctc tagggcagag ggccaccatc tcctgcaagt ccagccaaag tcttctgaat
      781 agtggtaatc agaagaacta cctcacctgg taccaacaga aaccaggaca gccacccaaa
      841 ctcctcatct attgggcatc cactagggaa tctgggatcc cagccaggtt tagtggcagt
      901 gggtctggga cagacttcac cctcaacatc catcctgtgg aggaggagga tgctgcaacc
      961 tattactgtc agaacgataa ttcccatccg ctcacgttcg gtgctgggac caagctggaa
     1021 ataaaaggcg atcctgtggc tcctaccgtg ctgatctttc caccagccgc tgatcaggtg
     1081 gccacaggca ctgtgacaat cgtgtgcgtg gccaacaagt acttccccga cgtgaccgtg
     1141 acctgggaag tcgatggcac cacacagacc accggcatcg agaacagcaa gacccctcag
     1201 aacagcgccg actgcaccta caacctgagc agcaccctga cactgaccag cacacagtac
     1261 aacagccaca aagagtacac ctgtaaagtc acccagggca ccaccagcgt ggtgcagagc
     1321 ttcaatagag gcgactgctg agcggcgcga ctctagatca taatcagcca taccacattt
     1381 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1441 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1501 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1561 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     1621 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1681 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1741 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1801 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1861 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1921 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1981 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2041 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2101 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2161 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2281 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2341 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2401 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2461 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2521 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2581 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2641 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2701 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     2761 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2821 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2881 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2941 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3001 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3061 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3121 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3181 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3241 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3301 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3361 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3421 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3481 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     3541 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3601 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3661 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3721 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3781 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     3841 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3901 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3961 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4021 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4081 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4141 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4201 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4261 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4321 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4381 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4441 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4501 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4561 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4621 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
