LOCUS       sequence-192595-        5732 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of secretory immunoglobulin heavy chains in mammalian 
            cells, human IgG4 isotype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  TITLE     Immunoglobulin expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5732)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AbVec2.0-IGHG4
FEATURES             Location/Qualifiers
     source          1..5732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    103..124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(139..144,145..162,163..169)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    177..193
                     /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     182..204
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     201..217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     201..217
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     826..850
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1060..1078
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1060..1077
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1091..1110
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2264..2273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2277..2411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2314..2333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2368..2387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2480..2837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2480..2671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2503..2523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2688..2823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2857..2874)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2857..2873)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2866..2888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3086..3541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3173..3192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3383..3404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3558..3577)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3677..3699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3737..3755)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3823..3927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3928..3996,3997..4788)
                     /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(4146..4165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4959..5547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5448..5467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5701..5718
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatcca actggcacga
       61 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      121 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      181 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt aattcgagct
      241 cgcccgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      841 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      901 ccgatccagc ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag
      961 tgacgtaagt accgcctata gagtctatag gcccaccccc ttggcttcgt tagaacgcgg
     1021 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     1081 aacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact gcacctcggt
     1141 tctatcgatt gaattccacc atgggatggt catgtatcat cctttttcta gtagcaactg
     1201 caaccggttg actaactagc tggtcgacca agggcccaag cgtcttcccc ctggcgccct
     1261 gctccaggag cacctccgag agcacagccg ccctgggctg cctggtcaag gactacttcc
     1321 ccgagccggt gacggtgtcg tggaactcag gcgccctgac cagcggcgtg cacaccttcc
     1381 cggctgtcct acagtcctca ggactctact ccctcagcag cgtggtgacc gtgccctcca
     1441 gcagcttggg cacgaagacc tacacctgca acgtagatca caagcccagc aacaccaagg
     1501 tggacaagag agttgagtcc aaatatggtc ccccatgccc atcatgccca gcacctgagt
     1561 tcctgggggg accatcagtc ttcctgttcc ccccaaaacc caaggacact ctcatgatct
     1621 cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccaggaagac cccgaggtcc
     1681 agttcaactg gtacgtggat ggcgtggagg tgcataatgc caagacaaag ccgcgggagg
     1741 agcagttcaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc
     1801 tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccgtcc tccatcgaga
     1861 aaaccatctc caaagccaaa gggcagcccc gagagccaca ggtgtacacc ctgcccccat
     1921 cccaggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctacc
     1981 ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca
     2041 cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaggcta accgtggaca
     2101 agagcaggtg gcaggagggg aatgtcttct catgctccgt gatgcatgag gctctgcaca
     2161 accactacac acagaagagc ctctccctgt ctctgggtaa atgagtgcca gggccggcaa
     2221 gcccccgctc cccgggctct cggggtcgcg cgaggaaagc ttggccgcca tggcccaact
     2281 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2341 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2401 atgtctggat cgggaattaa ttcggcgcag caccatggcc tgaaataacc tctgaaagag
     2461 gaacttggtt aggtaccttc tgaggcggaa agaaccatct gtggaatgtg tgtcagttag
     2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2761 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2821 gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt tacaacgtcg
     2881 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2941 cagttggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3001 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3061 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3301 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3421 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3481 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3541 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3601 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3661 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3721 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3781 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3901 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3961 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4021 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4081 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4141 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4201 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4261 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4321 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4381 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4441 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4501 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4561 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4621 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4681 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4741 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4801 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5581 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5641 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5701 agcgagtcag tgagcgagga agcggaagag cg
//
