LOCUS       sequence-503750-        8706 bp    DNA     circular SYN 26-MAR-2026
DEFINITION  Human IgG1 Fc protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8706)
  AUTHORS   Nettleship JE, Rada H, Owens RJ
  TITLE     Overview of a High-Throughput Pipeline for Streamlining the 
            Production of Recombinant Proteins.
  JOURNAL   Methods Mol Biol. 2019;2025:33-49. doi: 10.1007/978-1-4939-9624-7_2.
  PUBMED    31267447
REFERENCE   2  (bases 1 to 8706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 26, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2019"; volume: "2025:33-49. doi"; pages: " 
            10.1007/978-1-4939-9624-7_2"
COMMENT     Sequence Label: pOPINTTGneoFc
FEATURES             Location/Qualifiers
     source          1..8706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1239..1258
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1450..1471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1486..1491,1492..1509,1510..1516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1524..1540
                     /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     1548..1564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1560..1733
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(1580..1597)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1580..1596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1589..1611)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1753..1776
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2461..2478
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2553..2608
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2607..2626)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4844..4862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4930..5034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5035..5103,5104..5895)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5253..5272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6066..6654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6555..6574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6808..6825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6942..6963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6978..6983,6984..7001,7002..7008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7016..7032
                     /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     7040..7056
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7094..7114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7099..7428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7279..7414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7341..7360
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7495..8289
                     /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(7549..7568)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8159..8178
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8465..8586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8502..8521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8556..8575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
       61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
      781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
      841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
      901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
      961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     1261 ccactcccag gtccaagttt atactgaatt cttacaatca aaggagatat accatgggga
     1321 tccttcccag ccctgggatg cctgcgctgc tctccctcgt gagccttctc tccgtgctgc
     1381 tgatgggttg cgtagctgaa accggtaccg tcgaaccgac tggaaagcgg gcagtgagcg
     1441 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     1501 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     1561 tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1621 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1681 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     1741 aagctgttta aactggaagt tctgtttcag ggcccggaca aaactcacac atgcccaccg
     1801 tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc aaaacccaag
     1861 gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga cgtgagccac
     1921 gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca taatgccaag
     1981 acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt cctcaccgtc
     2041 ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa ctgggccctc
     2101 ccagccccca tcagcaaaac catctccaaa gccaaagggc agccccgaga accacaggtg
     2161 tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct gacctgcctg
     2221 gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg gcagccggag
     2281 aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt cctctacagc
     2341 aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg ctccgtgatg
     2401 cacgaggctc tgcacaacca ctacacgcag aagagcctct ccctgtctcc gggtaaaaaa
     2461 catcaccatc accatcacta agtgattaac ctcaggtgca ggcggccgct cgaggccggc
     2521 aaggccggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     2581 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2641 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     2701 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     2761 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     2821 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     2881 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     2941 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3001 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3061 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3121 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3181 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3241 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3301 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3361 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3421 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3481 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3541 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3601 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     3661 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     3721 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     3781 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     3841 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     3901 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     3961 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4021 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4081 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4141 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4201 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4261 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4321 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4381 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4441 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4501 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4561 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4621 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4681 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4741 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     4801 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6961 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7021 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7081 gaggtcgacc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     7141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     7201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     7261 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     7321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     7381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     7441 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     7501 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     7561 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     7621 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     7681 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     7741 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     7801 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     7861 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     7921 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     7981 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     8041 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     8101 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     8161 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     8221 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     8281 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     8341 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     8401 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     8461 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     8521 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     8581 atcttatcat gtctgtatac cgtcgacctc tagctagtcg accaattctc atgtttgaca
     8641 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     8701 tatgga
//
