LOCUS       Exported                4957 bp ds-DNA     circular SYN 23-APR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, May 7, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4957
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    85..372
                     /label=CAG enhancer
     misc_feature    805..1540
                     /label=B-actin promoter
     misc_feature    1716..1772
                     /label=DNA to be synthesized
     misc_feature    1716..1731
                     /label=16bp overhang for infusion cloning
     misc_feature    1733..1738
                     /label=Kozak
     misc_feature    1739..1759
                     /label=Secretion signal + scFv insertion site
                     /note="CD8a secretion signal: 5'- 
                     atggccttaccagtgaccgccttgctcctgccgctggccttgctgctccacgccgccag
                     gccg (amino acid: MALPVTALLLPLALLLHAARP)scFv linker: 5'- 
                     ggtggcggcggttccggaggtggtggttctggcggtggtggcagctcc (amino 
                     acid: GGGGSGGGGSGGGGSS)"
     misc_feature    1757..1772
                     /label=16bp overlap for infusion cloning
     CDS             1757..1759
                     /codon_start=1
                     /label=S
                     /translation="S"
     CDS             1760..1894
                     /codon_start=1
                     /locus_tag="CD8 hinge"
                     /label=CD8 hinge
                     /translation="TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD
                     "
     CDS             1895..1966
                     /codon_start=1
                     /locus_tag="CD8 TM"
                     /label=CD8 TM
                     /translation="IYIWAPLAGTCGVLLLSLVITLYC"
     misc_feature    2044..2100
                     /label=lac operator
     misc_feature    2044..2100
                     /label=beta-globin poly(A) sequence
     primer_bind     2460..2476
                     /label=M13 rev
     misc_feature    2508..2538
                     /label=lac promoter
     misc_feature    2553..2574
                     /label=CAP binding site
     misc_feature    2633..2828
                     /label=SV40 promoter
     misc_feature    2679..2814
                     /label=SV40 ori
     misc_feature    2834..2968
                     /label=SV40 poly(A)
     misc_feature    3207..3795
                     /label=ori
     misc_feature    3966..4826
                     /label=AMP
     misc_feature    4827..4931
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 ggccttaacc gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac
     1801 catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc
     1861 agtgcacacg agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg
     1921 gacttgtggg gttcttctcc tgtcactggt tatcaccctt tactgctaag ctagcagatc
     1981 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2041 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2101 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     2161 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     2221 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     2281 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     2341 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     2401 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     2461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     2641 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2701 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2761 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2821 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2941 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     3001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4921 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
