LOCUS       sequence-7123-ex        3642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3642)
  AUTHORS   Patterson GH, Lippincott-Schwartz J
  TITLE     A photoactivatable GFP for selective photolabeling of proteins and 
            cells.
  JOURNAL   Science. 2002 Sep 13. 297(5588):1873-7.
  PUBMED    12228718
REFERENCE   2  (bases 1 to 3642)
  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: "Science. 
            2002 Sep 13. 297(5588):1873-7."
COMMENT     Sequence Label: pRSETA-PAGFP
FEATURES             Location/Qualifiers
     source          1..3642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             68..90
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             110..127
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             131..163
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(167..175,176..190)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(206..208,209..211,212..922)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(251..272)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(512..531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     859..880
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(988..1006)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1002..1049
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1146..1601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1233..1252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1443..1464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1682..1786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1787..1855,1856..2647)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRPPRYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2818..3406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3307..3326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3560..3577
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaataatttt gtttaacttt aagaaggaga tatacatatg cggggttctc atcatcatca
      121 tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga
      181 tgacgataag gatcgatggg gatccatggt gagcaagggc gaggagctgt tcaccggggt
      241 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      301 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      361 caagctgccc gtgccctggc ccaccctcgt gaccaccttc agctacggcg tgcagtgctt
      421 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      481 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      541 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      601 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      661 tatcatggcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
      721 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
      781 ccccgtgctg ctgcccgaca accactacct gagccaccag tccaagctga gcaaagaccc
      841 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
      901 cggcatggac gagctgtaca agtaagaatt cgaagcttga tccggctgct aacaaagccc
      961 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1021 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg
     1081 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1141 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1321 atttagagct ttacggcacc tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag
     1381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atcgcggtct attcttttga
     1501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaat
     1561 atttaacgcg aattttaaca aaatattaac gtttacaatt tcgcctgatg cggtattttc
     1621 tccttacgca tctgtgcggt atttcacacc gcatacaggt ggcacttttc ggggaaatgt
     1681 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1801 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1861 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1921 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1981 aatgatgagc acttttaaag ttctgctatg tgatacacta ttatcccgta ttgacgccgg
     2041 gcaagagcaa ctcggtcgcc ccccgcggta ttctcagaat gacttggttg agtactcacc
     2101 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2161 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2221 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2281 ggagctgaat gaagccatac caaacgacga gagtgacacc acgatgcctg tagcaatgcc
     2341 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2401 aatagactga atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2461 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2521 agcactgggg ccagatggta agcgctcccg tatcgtagtt atctacacga cggggagtca
     2581 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2641 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2701 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2761 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2821 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2881 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2941 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3001 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3061 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3121 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3181 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3241 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3301 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3361 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3421 ggccttttta cggttcctgg gcttttgctg gccttttgct cacatgttct ttcctgcgtt
     3481 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3541 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3601 caaaccgcct ctccccgcgc gttggccgat tcattaatgc ag
//
