LOCUS       sequence-249990-        3646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the 3rd gen. 5' HIV LTR cassette as a type 8b part to be 
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3646)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK8b_004
FEATURES             Location/Qualifiers
     source          1..3646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1261..1386
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1879..2112
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2297..2341
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2608..2725
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     terminator      complement(2777..2811)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2882..3541)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3456..3475
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcacaata
      781 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
      841 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
      901 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
      961 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
     1021 cgatacaata aacggatctc tctggttaga ccagatctga gcctgggagc tctctggcta
     1081 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     1141 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     1201 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     1261 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     1321 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     1381 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     1441 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     1501 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     1561 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     1621 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     1681 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     1741 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     1801 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     1861 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     1921 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     1981 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     2041 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     2101 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     2161 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     2221 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     2281 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     2341 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     2401 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     2461 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     2521 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     2581 gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg gggggtacag
     2641 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     2701 acaaattaca aaaattcaaa attttatcga tcacgagact agcctcgaga agcttgatac
     2761 ccttgagacc agaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     2821 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     2881 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     2941 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     3001 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     3061 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgaaacg aaaaacatat
     3121 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     3181 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     3241 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     3301 caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt
     3361 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     3421 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     3481 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     3541 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     3601 ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaa
//
