LOCUS       Exported                8648 bp DNA     circular SYN 26-JAN-2022
DEFINITION  Membrane bound expression of human 41BBL, CD86 and HLA-A2 to make 
            aAPC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 25, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    6..303
                     /label=bGH-pA polyA signal
                     /note="bGH-pA"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #cc0000; direction: LEFT"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(314..355)
                     /note="5'-proxITR"
                     /note="modified region proximal to 5'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(361..585)
                     /label=5'-ITR
                     /note="5'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(586..589)
                     /note="tata"
                     /note="color: #cc0000"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3; direction: LEFT"
     promoter        705..809
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             810..1670
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla CDS
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:    1: 5567 .. 6358 / 
                     #ccffcc    2: 6359 .. 6427 / #ccffcc / signal sequence"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori rep origin
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of  
                     replication"
                     /note="color: #ffff00"
                     /note="color: #ffff00"
     misc_feature    complement(2613..2616)
                     /note="tata"
                     /note="color: #cc0000"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(2617..2841)
                     /label=3'-ITR
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(2842..2883)
                     /label=3'-proxITR
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(2894..3194)
                     /label=rBG-pA
                     /note="rBG-pA"
                     /note="rabbit beta-globin polyadenylation signal"
                     /note="color: #cc0000; direction: RIGHT"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(3212..3811)
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=pac from Streptomyces alboniger CDS
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /note="/Original 
                     Translation=MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
                     ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
                     QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
                     RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
                     /note="color: #993366; direction: LEFT"
     misc_feature    complement(3812..3877)
                     /label=P2A CDS
                     /note="Geneious type: Other"
                     /note="/created by=Sarah"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(3878..4864)
                     /label=Hu CD86
                     /note="/created by=Sarah"
                     /note="color: #993366; direction: LEFT"
     misc_feature    complement(4865..4930)
                     /label=P2A
                     /note="Geneious type: Other"
                     /note="/created by=Sarah"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(4931..5692)
                     /label=Hu 4-1BBL
                     /note="/created by=Sarah"
                     /note="color: #993366; direction: LEFT"
     misc_feature    5693..5698
                     /label=Kozak
                     /note="/created by=mclal85p"
                     /note="/modified by=Sarah"
                     /note="color: #a6acb3"
     promoter        complement(5710..6321)
                     /label=RPBSA promoter
                     /note="/created by=Sarah"
                     /note="color: #ffffff; direction: LEFT"
     exon            complement(5710..5719)
                     /label=RPL41 exon2
                     /note="RPL41 exon2"
                     /note="color: #cc0000; direction: RIGHT"
                     /note="color: #708090; direction: LEFT"
     intron          complement(5720..5834)
                     /label=RPL41 intron1
                     /note="/created by=Sarah"
                     /note="color: #------; direction: LEFT"
     exon            complement(5835..5887)
                     /label=RPL41 exon1
                     /note="/created by=Sarah"
                     /note="/modified by=Sarah"
                     /note="color: #708090; direction: LEFT"
     promoter        complement(5887..6321)
                     /label=RPL13a Promoter
                     /note="RPL13a Promoter"
                     /note="color: #cc0000; direction: RIGHT"
                     /note="color: #ffffff; direction: LEFT"
     promoter        6328..7520
                     /label=EF1a promoter
                     /note="EF1a promoter"
                     /note="constitutive promoter"
                     /note="color: #cc0000; direction: LEFT"
                     /note="color: #ffffff; direction: RIGHT"
     intron          6559..7501
                     /label=EF1a Intron1
                     /note="EF1a Intron1"
                     /note="color: #cc0000; direction: LEFT"
                     /note="color: #------; direction: RIGHT"
     exon            7502..7520
                     /label=EF1a Exon2
                     /note="EF1a Exon2"
                     /note="color: #cc0000; direction: LEFT"
                     /note="color: #708090; direction: RIGHT"
     misc_feature    7538..7542
                     /label=Kozak
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #00ee00; direction: RIGHT"
     gene            7543..8640
                     /label=HLA-A2
                     /note="/created by=Sarah"
                     /note="color: #a6acb3; direction: RIGHT"
ORIGIN
        1 aggccaagct tggggatcaa ttctctagag ctcgctgatc agcctcgact gtgccttcta
       61 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
      121 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
      181 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
      241 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
      301 cctactagat agcggacccc ttaccgaaac atcgccgcat tctgcagagg agtcgaactt
      361 agtgtatgta aacttctgac ccactggaat tgtgatacag tgaattataa gtgaaataat
      421 ctgtctgtaa acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac
      481 tgacttgcca aaactattgt ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt
      541 ttaatgactc caacttaagt gtatgtaaac ttccgacttc aactgtatag ggatccaatt
      601 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
      661 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2581 gcagcgagtc agtgagcgag gaagcggatc cctatacagt tgaagtcgga agtttacata
     2641 caccttagcc aaatacattt aaactcactt tttcacaatt cctgacattt aatcctagta
     2701 aaaattccct gtcttaggtc agttaggatc accactttat tttaagaatg tgaaatatca
     2761 gaataatagt agagagaatg attcatttca gcttttattt ctttcatcac attcccagtg
     2821 ggtcagaagt ttacatacac tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg
     2881 tcccgagtag ctagttcatg gcagccagca tatggcatat gttgccaaac tctaaaccaa
     2941 atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga gtgagagaca
     3001 caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc cagaagtcag
     3061 atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa gatctcagtg
     3121 gtatttgtga gccagggcat tggccacacc agccaccacc ttctgatagg cagcctgcac
     3181 ctgaggagtg aattcacgac aggccttcga atcaggcacc gggcttgcgg gtcatgcacc
     3241 aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt
     3301 agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg
     3361 ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg
     3421 atacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc
     3481 cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga
     3541 tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg
     3601 cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt
     3661 cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg
     3721 ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg
     3781 cgaggcgcac cgtgggcttg tactcggtca tggggccagg attctcctcg acgtcaccag
     3841 cctgcttcag caggctgaag ttagtagctc cggatccaaa acatgtatca cttttgtcgc
     3901 atgaagatgt cttcgaactt ttaaaaacac gctgggcttc atcagatctt tcaggtatat
     3961 ggattttttc tcttttcttg gtctgttcac tctcttccct ctccattgtg ttggttccac
     4021 atttataaga gttgcgaggc cgcttcttct tcttccattt ccatagaatt agacagaaaa
     4081 ccatcacaca tataataact gttggaagta cagctgtaat ccaaggaatg tggtctgggg
     4141 gaggctgagg gtcctcaagc tctatagaga aaggtgaaga taaaagccgc gtcttgtcag
     4201 tttccagaat acagaagatg gtcatattgc tcgtaacatc agggaatgaa acagacaagc
     4261 tgatggaaac gtcgtacagt tctgtgacat tatcttgaga tttctgcata ataccatcat
     4321 actcgatagt tgaattcttg gttcttagca aaacactcat cttcttaggt tctgggtaac
     4381 cgtgtataga tgagcaggtc aaatttatgt acacattttc tgttatatta gaaattggta
     4441 ctatttcagg ttgactgaag ttagcaagca ctgacagttc agaattcatc tggtggatgc
     4501 gaatcattcc tgtgggcttt ttgtgatgga tgatacattg atacaagccc ttgtccttga
     4561 tctgaagatt gtgaagtctc agggtccaac tgtccgaatc aaaacttgtg cggcccatat
     4621 acttggaatg aacactgtca aatttctctt tgcctaagta tacctcattc agaaccaagt
     4681 tttcctggtc ctgccaaaat actactagct cactcaggct ttggttttga gagtttgcaa
     4741 attggcatgg caggtctgca gtctcattga aataagcttg aatcttcaga ggagcagcac
     4801 cagagagcag gaaggccatc acaaagagaa tgttactcag tcccatagtg cactggggat
     4861 ccatggggcc aggattctcc tcgacgtcac cagcctgctt cagcaggctg aagttagtag
     4921 ctccggatcc ttccgacctc ggtgaaggga gtccggctgg gatttcgggg gtcacccgga
     4981 agagtcccaa gactgtggcg ccctgggtaa gctgccaggc atggcgtgcc ctggcctcag
     5041 tgtgaagatg gacgcccagg cgctggccgg cactcaggtg cagcaagcgg ccctggaaac
     5101 cgaaggccga gttccgagcc tcggaggagg cgggtggcag gtccacggtc aaagccaggg
     5161 cggcggcccc agcagcagag cgcagtggct gcaggtgcag cgcaagtgaa acggagcctg
     5221 agccctcgcc ggccaccacg cgccgcagct ctagttgaaa gaagacatag tagactccag
     5281 ccttggccac caccagctcc ttcgtgtcct ctttgtagct caggcccccc gtcagggaca
     5341 cgcctgccag gcctgggtca ctgtaccagc tcaggggccc atcgatcagc agaacatttt
     5401 gggccaccag ctgcgcaaac atgccctgcc gcaggtccaa gaggccggcg ggatcgtcgg
     5461 gcgaaagctc gggaccctcg cggagtctcg ggctggccgc ggagccgggc gaggcgcgag
     5521 ccccggacac ggcccagggg caggcgagga agacggcgca ggcggcagcg agcagcagca
     5581 gcagcagcag ccccgcgacc agggcccaag gcagtacgcg gcaggcgcga gcgcggggcg
     5641 cgggaggcca cggggcttcg gggtccagtg aagcgtcaga ggcgtattcc atggtggccg
     5701 ccctcaggtg cagaggtttc tacagggaaa gagagaaagg ctacagttag tttgctgtga
     5761 tgatgctaag tgccgaggtc agatgtctat ggattgtaca cggaatccca acgcaggaac
     5821 caaaaaacac tcacccaaaa aaatggcgct aaggccgaga gaaaggcgaa gttccgtcta
     5881 cggctatata ccttgaaaca ggggcggcag aagtgacgca acgcggttgc gcggacggct
     5941 tcctgtgttt tcattggtcg aagcgttctg cgcgtgcgca gagattcccc agggaaggcg
     6001 ggccctcttc cgaatgtcca atccggacat tcttccggtt ggacctccgc ggtcagcgct
     6061 gtgcccggcg agagatcacg tggggcgcgg aggcggtgct gctggggcac ggccgtccag
     6121 cctcggcggc catatttttg aggggctgtt catctcgttc acacgctctg tccgccatgt
     6181 ttgtgagtgg aagcgccatt accccttcaa gcgactgaag gctgcagggc ctctggtggc
     6241 ccgcatgggg agaccagacc cgccaggccc gcctttccgc actcagtccg ggcttacttt
     6301 attttgtgag acagggtctc gcctagaggc tccggtgccc gtcagtgggc agagcgcaca
     6361 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     6421 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     6481 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     6541 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
     6601 ggttatggcc cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct
     6661 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     6721 cccttcgcct cgtgcttgag ttgaggcttg gcctgggcgc tggggccgcc gcgtgcgaat
     6781 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     6841 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     6901 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     6961 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     7021 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     7081 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     7141 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     7201 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     7261 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     7321 tttaggttgg ggggaggggt tttatgcgat ggagtttccc catactgagt gggtggagac
     7381 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     7441 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttaa
     7501 ggtgtcgtga aaactacccc aagctggcct ctgaggccca ccatggccgt catggcgccc
     7561 cgaaccctcg tcctgctact ctcgggggct ctggccctga cccagacctg ggcgggctct
     7621 cactccatga ggtatttctt cacatccgtg tcccggcccg gccgcgggga gccccgcttc
     7681 atcgcagtgg gctacgtgga cgacacgcag ttcgtgcggt tcgacagcga cgccgcgagc
     7741 cagaggatgg agccgcgggc gccgtggata gagcaggagg gtccggagta ttgggacggg
     7801 gagacacgga aagtgaaggc ccactcacag actcaccgag tggacctggg gaccctgcgc
     7861 ggctactaca accagagcga ggccggttct cacaccgtcc agaggatgta tggctgcgac
     7921 gtggggtcgg actggcgctt cctccgcggg taccaccagt acgcctacga cggcaaggat
     7981 tacatcgccc tgaaagagga cctgcgctct tggaccgcgg cggacatggc agctcagacc
     8041 accaagcaca agtgggaggc ggcccatgtg gcggagcagt tgagagccta cctggagggc
     8101 acgtgcgtgg agtggctccg cagatacctg gagaacggga aggagacgct gcagcgcacg
     8161 gacgccccca aaacgcatat gactcaccac gctgtctctg accatgaagc caccctgagg
     8221 tgctgggccc tgagcttcta ccctgcggag atcacactga cctggcagcg ggatggggag
     8281 gaccagaccc aggacacgga gctcgtggag accaggcctg caggggatgg aaccttccag
     8341 aagtgggcgg ctgtggtggt gccttctgga caggagcaga gatacacctg ccatgtgcag
     8401 catgagggtt tgcccaagcc cctcaccctg agatgggagc cgtcttccca gcccaccatc
     8461 cccatcgtgg gcatcattgc tggcctggtt ctctttggag ctgtgatcac tggagctgtg
     8521 gtcgctgctg tgatgtggag gaggaagagc tcagatagaa aaggagggag ctactctcag
     8581 gctgcaagca gtgacagtgc ccagggctct gatgtgtctc tcacagcttg taaagtgtga
     8641 ggcctgtc
//
