LOCUS       sequence-360595-        6715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of TH rat mAb LC gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6715)
  AUTHORS   Huang KL, Zhu H
  TITLE     Heng Zhu lab antibody-expressing plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6715)
  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            "unpublished"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_LC_Rat Ig Kappa
FEATURES             Location/Qualifiers
     source          1..6715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        447..650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     601..621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     647..671
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    1508..2096
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1561..1581)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1979..1990)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2118..2137)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2162..2210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2412..2840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2499..2518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2709..2730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2849..2869)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2854..3183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3034..3169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3096..3115
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3250..4044
                     /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(3304..3323)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3914..3933
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4220..4341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4257..4276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4311..4330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4390..4406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4390..4406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4403..4425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4414..4430
                     /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(4438..4444,4445..4462,4463..4468))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4483..4504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4621..4638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4872..4891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5551..6342,6343..6411))
                     /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     6174..6193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6412..6516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6591..6610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
       61 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      121 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      421 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      481 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      541 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      601 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      661 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
      721 accgatccag cctccggact ctagagaatt cgccaccatg agtcctgccc agttcctgtt
      781 tctgctagtg ctctcgattc aggaaaccaa cggtgatgtt gtgatgactc agaccccact
      841 cactttgtcg gttaccattg gacaaccagc ctccatctct tgcaaatcaa gtcagagcct
      901 cttacatagt aatggaaaga catatttgaa ttggttatta cagaggccag gccagtctcc
      961 aaagctccta atctatctgg tgtctaaact ggaatctgga gtccctgaca ggttcagtgg
     1021 cagtggatca gggacagatt tcacactgaa aatcagcaga gtggaggctg aggatttggg
     1081 agtttattac tgcttgcaag ctacacattt tcctcagacg ttcggtggag gcaccaagct
     1141 ggaaatcaaa gccgacgccg ccccaaccgt gtctatcttc ccccctagca tggagcagct
     1201 gacctctggc ggcgccacag tggtgtgctt cgtgaacaac ttctaccccc gggacatctc
     1261 cgtgaagtgg aagatcgatg gctctgagca gagagacggc gtgctggata gcgtgaccga
     1321 ccaggattct aaggatagca catattccat gagctccacc ctgtccctga caaaggtgga
     1381 gtacgagagg cacaacctgt atacctgcga ggtggtgcac aagacatcta gctcccccgt
     1441 ggtgaagtcc tttaaccgga atgagtgttg aaagcttacc ggttagtaat gagtttgata
     1501 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1561 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1621 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1681 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1741 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1801 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1861 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     1921 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     1981 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2041 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     2101 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     2161 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     2221 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     2281 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     2341 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     2401 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2461 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2521 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     2581 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2641 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2701 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2761 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     2821 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3421 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     4081 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4141 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4201 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4261 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4321 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4381 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4441 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4501 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4561 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4621 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4681 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5221 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5581 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5641 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5701 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5761 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5821 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5881 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5941 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6001 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6061 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6121 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6181 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6241 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6301 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6361 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6541 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     6601 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6661 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgca
//
