LOCUS       sequence-381530-        5353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CoVMab4 light chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5353)
  TITLE     Murrell lab nanobodies
REFERENCE   2  (bases 1 to 5353)
  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"
COMMENT     Sequence Label: hIgG1_kappa_CoVMab4
FEATURES             Location/Qualifiers
     source          1..5353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     20..37
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     51..70
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      845..854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    858..992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(895..914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     949..968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1061..1418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1061..1252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1084..1104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1269..1404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1438..1455)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1438..1454)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1447..1469)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1667..2122
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1754..1773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1964..1985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2139..2158)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2258..2280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2318..2336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2404..2508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2509..2577,2578..3369)
                     /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     complement(2727..2746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3540..4128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4029..4048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4282..4299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4416..4437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4452..4457,4458..4475,4476..4482)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4490..4506
                     /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)."
     primer_bind     4495..4517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4514..4530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4514..4530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        4559..4938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4939..5142
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5093..5113
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5139..5163
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tatgtatcat acacatacga tttaggtgac actatagaat aacatccact ttgcctttct
       61 ctccacaggt gtccactccc aggtccaact gcacctcggt tctatcgatt gaattccacc
      121 atgggatggt catgtatcat cctttttcta gtagcaactg caaccggtgt acactcggat
      181 gttttgatga cccaaactcc actctccctg cctgtcagtc ttggagatca agcctccatc
      241 tcttgcagat ctagtcagac cattgtacat actggtggaa acacctattt agattggtac
      301 ctgcagaaac caggccagtc tccaaaactc ctgatttaca aagtttccaa ccgattttct
      361 ggggtcccag acaggttcag tggcagtgga tcagggacag atttcacact caggatcagc
      421 agagtggagg ctgaggatct gggagtttat ttctgctttc aaggttcaca tgttccattc
      481 acgttcggct cggggacaaa gttggaaata aaacgtacgg tggctgcacc atctgtcttc
      541 atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg
      601 aataacttct atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcg
      661 ggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc
      721 agcaccctga cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc
      781 acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttagaag
      841 cttggccgcc atggcccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
      901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
      961 aactcatcaa tgtatcttat catgtctgga tcgggaatta attcggcgca gcaccatggc
     1021 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccatc
     1081 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     1141 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     1201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     1261 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     1321 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     1381 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctgttaaca gcttggcact
     1441 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1501 tgcagcacat ccccctttcg ccagttggcg taatagcgaa gaggcccgca ccgatcgccc
     1561 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     1621 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     1681 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1741 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1801 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1861 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     1921 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1981 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     2041 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2101 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     2161 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2221 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2281 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2341 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3661 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat ccaactggca cgacaggttt
     4381 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4441 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4501 taacaatttc acacaggaaa cagctatgac atgattacga attaattcga gctcgcccga
     4561 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4621 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4681 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4741 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4801 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4861 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4921 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4981 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     5041 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     5101 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     5161 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     5221 agcctccgcg gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta
     5281 agtaccgcct atagagtcta taggcccacc cccttggctt cgttagaacg cggctacaat
     5341 taatacataa cct
//
