LOCUS       sequence-79877-e        7349 bp    DNA     circular SYN 14-APR-2025
DEFINITION  jxOFF-post.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7349)
  TITLE     Cre-dependent OFF post-mGRASP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7349)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: paavCAG-JxOFF-post-mGRASP-2A-dTomato
FEATURES             Location/Qualifiers
     source          1..7349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        701..978
                     /label=chicken beta-actin promoter
     primer_bind     1258..1280
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1342..1361
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1393..1426)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2503..2514)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2911..2964
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    3697..3730
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3743..4331
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3796..3816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4214..4225)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4374..4495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4385..4404)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4439..4458
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4657..4786
                     /label=AAV2 ITR
     primer_bind     complement(4805..4822)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4805..4821)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4814..4836)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4963..5418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5050..5069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5260..5281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5444..5548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5549..5617,5618..6409)
                     /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"
     primer_bind     complement(5767..5786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6580..7168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7069..7088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatcgta ccattgacgt caataatgac gtatgttccc
      481 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact
      541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      661 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      721 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      781 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      841 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      901 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      961 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc
     1021 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1081 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     1141 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     1201 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1261 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1321 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1381 aagaattgga tcataacttc gtatagcata cattatacga acggtaaagc ttgatgatcc
     1441 atggcacttc ctagatgtat gtggcctaat tatgtatggc gagctatgat ggcctgcgtg
     1501 gtgcaccgag ggagtggcgc cccacttaca ctctgcctgt tgggttgcct cctccagact
     1561 ttccacgttc tttcacagaa gctggatgac atgtccaagg gagaagagtt gtttactggc
     1621 gtagtcccta ttctcgtgga actcgatggt gacgtgaatg gccataagtt ttctgtcaga
     1681 ggagagggag aaggcgatgc caccatcggg aaactcacgc tgaaattcat ctgtaccact
     1741 ggaaaacttc ccgtgccttg gccaaccctc gtgacaacac tcacctacgg ggtgcaatgt
     1801 ttctctcggt acccggacca tatgaagagg catgacttct tcaagagtgc catgcccgag
     1861 ggttacgttc aggagcgcac catctctttt aaggacgatg gcaaatataa gacaagagca
     1921 gtcgtcaagt tcgagggtga tacactcgtt aaccgcatcg agctcaaagg caccgatttt
     1981 aaggaggacg gaaatatcct gggacacaaa ttggagtaca acttcaacag tcacaacgtg
     2041 tatattacag cagataagca gaagaatggc ataaaggcca atttcacggt aagacataat
     2101 gtcgaggatg gcagtgtcca gctggcagac cactaccagc aaaacacgcc cattggcgat
     2161 ggacctgttc tcctgccaga caaccactac ctcagtaccc aaacagtcct gtccaaggac
     2221 cctaatgaga aagggacgct cgagctggtg ccccacttgc ataacctcaa tgacatcagc
     2281 caatacacaa gtacaaccac caaggtccca agcactgaca tcacccttcg accaactcgg
     2341 aaaaactcaa cccctgtgac cagcgcattt ccaaccgcta agcaagacga ccccaaacaa
     2401 cagcctagtc catttagcgt agaccaaagg gactacagca cagagctgtc cgtgaccata
     2461 gctgtaggag cttccctgct tttcctgaac attctggctt tcgcggccct ctattacaag
     2521 aaggacaaaa ggcgccacga cgtacatagg agatgtagtc cgcaacgaac tacaaccaat
     2581 gatcttacgc acgcacctga ggaggagatc atgagtcttc agatgaaaca cacagacctg
     2641 gaccacgagt gtgaatccat ccatccccat gaggtggtgc tgaggacagc atgccctccg
     2701 gactataccc tggcaatgcg ccgaagccca gacgacattc ccctgatgac acccaatacc
     2761 ataaccatga ttcccaacac tattccagga atccagcctc tccacacttt caacacattc
     2821 actggtggac agaataatac tctccctcat ccgcatccac acccccactc ccacagcaca
     2881 acacgggtgg gtggcagcgg tggcagatct gagggtcggg gctctctgct cacatgtggc
     2941 gacgtcgagg agaatcccgg accggccccg gggtcgacaa tggtgtctaa gggtgaggaa
     3001 gtaattaagg agtttatgag attcaaagtt agaatggagg gtagcatgaa cgggcacgag
     3061 tttgaaattg aaggtgaggg tgaaggaaga ccatatgagg gcacacaaac cgccaagctc
     3121 aaggttacca aaggtggccc ccttcctttc gcctgggaca tcctcagccc tcaattcatg
     3181 tacggtagca aggcttacgt caaacaccct gctgatatac ctgattacaa aaaactgtca
     3241 tttcctgaag gatttaagtg ggaacgagta atgaattttg aggatggtgg gctggtcact
     3301 gtcacgcaag actcctctct gcaggacggc accctgattt acaaggtgaa gatgcgcgga
     3361 acaaattttc cccccgatgg ccccgtgatg cagaagaaga ccatggggtg ggaggcctct
     3421 acagaacgac tgtacccccg cgatggcgtt cttaaaggcg aaatccatca ggcgcttaaa
     3481 ctcaaagatg gcgggcacta cctggttgag ttcaaaacta tatatatggc caaaaagccg
     3541 gttcagctgc ctggctacta ctacgtggac accaagttgg acatcacgag tcacaacgag
     3601 gactacacaa tcgtcgagca gtacgaacgc tcagaaggtc ggcatcacct gttcttgtat
     3661 ggcatggatg agctgtataa gtaagctatc agatctataa cttcgtataa tgtatgctat
     3721 acgaacggta cagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3781 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3841 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3901 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3961 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4021 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4081 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4141 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4201 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4261 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4321 cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata
     4381 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4441 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4501 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     4561 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     4621 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     4681 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4741 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
     4801 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     4861 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     4921 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4981 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5041 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5101 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5161 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5221 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5281 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5341 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5401 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5461 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5521 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5581 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5641 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5701 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5761 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5821 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5881 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5941 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6001 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6061 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6121 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6181 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6241 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6301 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6361 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6421 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6481 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7201 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7261 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7321 cagcgagtca gtgagcgagg aagcggaag
//
