LOCUS       sequence-211137-        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NpHR-YFP reporter vector of iTango and Cal-Light system for AAV 
            production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  TITLE     fluorescent protein reporter vectors of iTango and Cal-Light system 
            for AAV production
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6028)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-TRE-NpHR-YFP
FEATURES             Location/Qualifiers
     source          1..6028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    join(298..316,317..339,340..358,359..381,382..400,401..423,
                     424..442,443..465,466..484,485..507,508..526,527..549,
                     550..568)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..639
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(1693..1695,1696..2406)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1735..1756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1996..2015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2343..2364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2443..3031
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2496..2516)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2914..2925)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3066..3083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3072..3279
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3302..3431
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3493..4081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3573..3592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4252..5043,5044..5112))
                     /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     4875..4894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5113..5217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5285..5303
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5341..5363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5463..5482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5499..5954)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5636..5657)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5848..5867
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
      661 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      721 ctccgcggcc ccgaattcga gctcggtacc cggggatcca tgaggggtac gcccctgctc
      781 ctcgtcgtct ctctgttctc tctgcttcag gacacagaga ccctgcctcc cgtgaccgag
      841 agtgccgtgg cccttcaagc cgaggttacc caaagggagt tgttcgagtt cgtgctgaac
      901 gaccctttgc ttgcaagcag tctctatatc aacatcgcac ttgcaggact gagtatactg
      961 ctgttcgttt ttatgacccg aggactcgat gatccacggg caaaacttat tgctgtgtca
     1021 accatccttg tgcctgtcgt cagcattgcc tcctacactg gattggcgag cggcctgaca
     1081 atttccgttc ttgaaatgcc agcgggccat tttgcagaag gcagctcagt gatgctggga
     1141 ggagaagagg tagatggtgt agtcaccatg tggggacggt atctcacctg ggcactttcc
     1201 acgcccatga ttctcctcgc tctgggtctc ctggccggaa gcaatgctac aaagctcttc
     1261 acagctatca ctttcgatat cgctatgtgc gtgactggcc ttgccgcggc cctgactacc
     1321 tcctcccacc tcatgagatg gttctggtac gctatcagtt gtgcatgctt tctggtggtc
     1381 ttgtatatcc tgctggtgga gtgggcacag gacgccaaag ccgcgggaac cgctgacatg
     1441 ttcaataccc tgaagctgtt gacagtagtg atgtggctgg ggtatccaat tgtgtgggct
     1501 cttggagtcg agggtatcgc ggtgttgccc gttggggtga cgagctgggg atattctttc
     1561 ctggatatcg tggcaaagta cattttcgca ttcttgctcc tgaactatct gacgtcaaac
     1621 gaatctgtcg tgtccggcag cattttggat gttccatctg cttctgggac cccggctgat
     1681 gatgcggccg ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1741 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1801 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1861 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
     1921 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1981 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2041 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2101 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2161 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2221 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2281 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2341 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2401 tacaagttct gctacgagaa cgaggtgtaa aagcttatcg ataatcaacc tctggattac
     2461 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2521 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2581 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2641 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2701 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2761 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2821 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     2881 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2941 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3001 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatcg
     3061 atctgcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3241 gggggaggat tgggaagaca atagcaggca tgctggggac acgtgcggac cgagcggccg
     3301 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3361 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3421 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3481 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3541 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3601 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3661 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3721 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3781 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3841 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3901 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3961 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4021 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4081 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4141 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4201 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4261 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4321 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4381 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4441 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4501 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4561 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4621 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4681 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4741 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4801 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4861 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4921 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4981 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5041 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5101 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5161 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5221 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5281 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5341 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5401 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5461 tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt
     5521 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat
     5581 cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt
     5641 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5701 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     5761 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5821 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc
     5881 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5941 gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc
     6001 gtaaggagaa aataccgcat caggcgcc
//
