LOCUS       sequence-431737-        6966 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Generate lentiviruses to act as a positive transduction control for 
            pHAGE2-TetO vectors in pluripotency reprogramming.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  TITLE     Jauch lab plasmids
REFERENCE   2  (bases 1 to 6966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pHAGE2-TetO-mCherry
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(2..20,21..43,44..62,63..85,86..104,105..127,128..146,
                     147..169,170..188,189..211,212..230,231..253,254..272)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     340..364
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             435..1145
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(580..598)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     848..867
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1159..1747
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1212..1232)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1630..1641)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1895..2075
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2122..2140)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2208..2312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2313..2381,2382..3173)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2531..2550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3344..3932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4086..4103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4343..4364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4379..4384,4385..4402,4403..4409)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4417..4433
                     /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     4422..4444
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4441..4457
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4441..4457
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4507..4526
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4718..5351
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5398..5523
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6020..6253
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6438..6482
                     /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    6775..6892
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
       61 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
      121 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      181 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      241 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagctc ggtacccggg
      301 tcgaggtagg catgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca
      361 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      421 cagcctgcgg ccgcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
      481 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
      541 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
      601 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
      661 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
      721 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
      781 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
      841 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
      901 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
      961 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1021 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1081 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1141 agtaaatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1201 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1261 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1321 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1381 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     1441 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1501 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     1561 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     1621 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     1681 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     1741 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     1801 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatcacct
     1861 gcaggacagg cgcgccctgc tttttgcttg tactgggtct ctctggttag accagatctg
     1921 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     1981 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     2041 cagacccttt tagtcagtgt ggaaaatctc tagcacccgg gcgattaagg aaagggctag
     2101 atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2161 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2281 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2401 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2521 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2581 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2701 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3961 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4021 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4081 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4141 ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt tttttattta
     4201 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4261 tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag
     4321 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4381 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4441 caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt
     4501 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     4561 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     4621 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa
     4681 attgtatttg ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa
     4741 agaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag
     4801 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag
     4861 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg
     4921 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg
     4981 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag
     5041 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt
     5101 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
     5161 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5221 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5281 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5341 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     5401 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     5461 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     5521 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     5581 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     5641 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     5701 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5761 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5821 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca
     5881 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     5941 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6001 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     6061 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     6121 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     6181 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     6241 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     6301 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     6361 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     6421 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     6481 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     6541 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     6601 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     6661 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     6721 tgaacggatc tcgacggtat cgccgaattc acaaatggca gtattcatcc acaattttaa
     6781 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     6841 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     6901 ttacagggac agcagagatc cagtttggac tagtccacac cctaactgac acactcgagt
     6961 ttacca
//
