LOCUS       pTwist_CMV_mIgG2        7344 bp    DNA     circular SYN 04-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..7344,1..1174)
                     /organism="pTwistCMVmIgG2a(truncated) in pTwist CMV BG WPRE
                     Neo v2.0"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          642..1117
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     misc_feature    1174^1175
                     /label=Your gene here
     misc_feature    1175..1885
                     /label=VHH mIgG2a
     misc_feature    1910..2498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2381..2392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2551..2672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2787..2833
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2943..3014
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3059..3255
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3106..3241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3269..4063
                     /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"
     polyA_signal    4074..4298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4494..5082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5828..5932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5933..6001,6002..6793)
                     /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"
     terminator      6806..6942
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7087..7158
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    7216..7264
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tcccctcgaa
      601 gcttacatgt ggtaccgagc tcggatcctg agaacttcag ggtgagtcta tgggaccctt
      661 gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag gggagaagta
      721 acagggtaca catattgacc aaatcagggt aattttgcat ttgtaatttt aaaaaatgct
      781 ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc
      841 tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag
      901 tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa
      961 attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc
     1021 tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt
     1081 gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg
     1141 tgtgctggcc catcactttg gcaaaggcgg ccgcggcgga tccgagccta gaggccctac
     1201 catcaaaccc tgtcctccat gcaagtgccc cgctcctaat ctgctcggag gccccagcgt
     1261 gttcatcttc ccacctaaga tcaaggacgt gctgatgatc tctctgagcc ccatcgtgac
     1321 ctgcgtggtg gtggatgtgt ccgaggacga tcccgatgtg cagatcagtt ggttcgtgaa
     1381 caacgtggaa gtgcacacag cccagacaca gacccacaga gaggactaca acagcaccct
     1441 gagagtggtg tctgccctgc ctatccagca ccaggattgg atgagcggca aagaattcaa
     1501 gtgcaaagtg aacaacaagg acctgcctgc tcctatcgag cggaccatct ctaagcctaa
     1561 gggctctgtt agagcccctc aggtgtacgt gctgcctcct ccagaggaag agatgaccaa
     1621 gaaacaagtg accctgacct gtatggtcac cgacttcatg cccgaggaca tctacgtgga
     1681 atggaccaac aacggcaaga ccgagctgaa ctacaagaac acagagcccg tgctggacag
     1741 cgacggcagc tacttcatgt acagcaagct gcgcgtggaa aagaagaact gggtcgagcg
     1801 gaacagctac agctgctctg tggtgcacga gggcctgcac aatcaccaca ccaccaagag
     1861 cttcagcaga accccaggca agtgaatcga tttctggatc cgcaggccta atcaacctct
     1921 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1981 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2041 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2101 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2161 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2221 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2281 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2341 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2401 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2461 tcagacgagt cggatctccc tttgggccgc ctccccgcct gtctagcttg actgactgag
     2521 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     2581 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2641 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2701 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2761 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     2821 gaggggtttt ttgtgcccct cgggccggat tgctatctac cgaggcactg gtgtactgat
     2881 agagccgggc tagatgttaa ctgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     2941 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     3001 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aagcgatctg
     3061 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3121 ccgcccagtt ccgcccattc tccgccccat cgctgactaa ttttttttat ttatgcagag
     3181 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3241 ctaggctttt gcaaaggagg tagccaacat gattgaacaa gatggattgc acgcaggttc
     3301 tcccgccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3361 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3421 cgacctgtcc ggtgccctga atgaactcca ggacgaggca gcgcggctat cgtggctggc
     3481 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3541 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3601 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3661 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3721 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3781 cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc acggcgatgc
     3841 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3901 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3961 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4021 gcagcgcatc gccttctatc gccttcttga cgagttcttc tagtatgtaa gccctgtgcc
     4081 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4141 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4201 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4261 caatagcagg catgctgggg atgcggtggg ctctatggtt aattaaccag tcaagtcagc
     4321 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     4381 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5281 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     5341 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     5401 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     5461 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     5521 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     5581 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     5641 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     5701 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     5761 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     5821 ctttcgccgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     6061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6121 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6241 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     6361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6481 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     6541 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6781 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     6841 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     6901 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt aaactcgact
     6961 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     7021 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     7081 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     7141 cagcttcagc tacctctcaa tttgcgtata gagcgggtca tcgaagggtt atacactcgt
     7201 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     7261 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     7321 tcatagccca tatatggagt tccg
//
