LOCUS       sequence-380456-        4666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  alpha-Tubulin light chain variable region + rabbit light chain 
            constant region + exogenous N-term signal peptide. For co-expression
            with pDL023 in HEK293 cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4666)
  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 4666)
  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: pDL024_Tub-rb_IgG_LC
FEATURES             Location/Qualifiers
     source          1..4666
                     /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    1454..1575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1491..1510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1545..1564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1582..2037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1719..1740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1931..1950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2064..2168
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2170..2527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2170..2361
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2193..2213)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2378..2513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2562..3356
                     /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             2562..3356
                     /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(2616..2635)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3226..3245
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3544..3563)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3588..3635
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3964..4552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4453..4472
                     /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 cagatgtcaa agcgtgctga cccagcctcc tagcgcctct
      721 ggaacacctg gacagagagt gaccatcagc tgtagcggca gcagcagcaa catcggcagc
      781 aactacgtgt actggtatca gcagctgccc ggcacagccc ctaaactgct gatctaccgg
      841 aacaaccagc ggcctagcgg cgtgccagat agattttctg gcagcaagag cggcacctct
      901 gccagcctgg ctatctctgg actgagaagc gaggacgagg ccgactacta ttgtgccgcc
      961 tgggatgata gcctggccag cgttgtgttt ggcggaggca caaagctgac cgtgcttggc
     1021 gatcctgtgg ctcctaccgt gctgatcttt ccaccagccg ctgatcaggt ggccacaggc
     1081 actgtgacaa tcgtgtgcgt ggccaacaag tacttccccg acgtgaccgt gacctgggaa
     1141 gtcgatggca ccacacagac caccggcatc gagaacagca agacccctca gaacagcgcc
     1201 gactgcacct acaacctgag cagcaccctg acactgacca gcacacagta caacagccac
     1261 aaagagtaca cctgtaaagt cacccagggc accaccagcg tggtgcagag cttcaataga
     1321 ggcgactgct gagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1381 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1441 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1561 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1621 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1681 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1741 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1801 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1861 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1921 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1981 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2161 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2221 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2281 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2341 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2401 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2461 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2521 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2581 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2641 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2701 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2761 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2821 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2881 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2941 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3001 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3061 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3121 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3181 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3241 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3301 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3361 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3421 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3481 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     3541 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3601 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3661 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3721 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3781 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     3841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3901 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3961 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4021 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4081 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4141 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4201 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4261 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4321 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4381 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4441 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4501 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4561 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4621 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
