LOCUS       sequence-240470-        6181 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 6181)
  AUTHORS   Zhuo L, Sun B, Zhang CL, Fine A, Chiu SY, Messing A
  TITLE     Live astrocytes visualized by green fluorescent protein in 
            transgenic mice.
  JOURNAL   Dev Biol. 1997 Jul 1;187(1):36-42.
  PUBMED    9224672
REFERENCE   2  (bases 1 to 6181)
  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: "Dev Biol.";
            date: "1997-07-1"; volume: "187(1)"; pages: "36-42"
COMMENT     Sequence Label: hGFAP-GFP
FEATURES             Location/Qualifiers
     source          1..6181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /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"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1626..1645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1879..1896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2013..2034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2049..2054,2055..2072,2073..2079)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2087..2103
                     /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     2092..2114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2111..2127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2111..2127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      4382..4391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4388..4390,4391..4393,4394..5107)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLM
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4433..4454)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4694..4713)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5692..5709)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5692..5708)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5701..5723)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5917..5936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6036..6058
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6096..6114)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1981 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2101 caatttcaca caggaaacag ctatgacatg attacgaatt cgagctcggt accagatctg
     2161 agctcccacc tccctctctg tgctgggact cacagaggga gacctcagga ggcagtctgt
     2221 ccatcacatg tccaaatgca gagcataccc tgggctgggc gcagtggcgc acaactgtaa
     2281 ttccagcact ttgggaggct gatgtggaag gatcacttga gcccagaagt tctagaccag
     2341 cctgggcaac atggcaagac cctatctcta caaaaaaagt taaaaaatca gccacgtgtg
     2401 gtgacacaca cctgtagtcc cagctattca ggaggctgag gtgaggggat cacttaaggc
     2461 tgggaggttg aggctgcagt gagtcgtggt tgcgccactg cactccagcc tgggcaacag
     2521 tgagaccctg tctcaaaaga caaaaaaaaa aaaaaaaaaa aaaagaacat atcctggtgt
     2581 ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg
     2641 gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc
     2701 ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg
     2761 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa
     2821 gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa
     2881 aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca
     2941 aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg
     3001 ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt
     3061 gaggggactg ggcagggttc tgaccctgtg ggaccagagt ggagggcgta gatggacctg
     3121 aagtctccag ggacaacagg gcccaggtct caggctccta gttgggccca gtggctccag
     3181 cgtttccaaa cccatccatc cccagaggtt cttcccatct ctccaggctg atgtgtggga
     3241 actcgaggaa ataaatctcc agtgggagac ggaggggtgg ccagggaaac ggggcgctgc
     3301 aggaataaag acgagccagc acagccagct catgtgtaac ggctttgtgg agctgtcaag
     3361 gcctggtctc tgggagagag gcacagggag gccagacaag gaaggggtga cctggaggga
     3421 cagatccagg ggctaaagtc ctgataaggc aagagagtgc cggccccctc ttgccctatc
     3481 aggacctcca ctgccacata gaggccatga ttgaccctta gacaaagggc tggtgtccaa
     3541 tcccagcccc cagccccaga actccaggga atgaatgggc agagagcagg aatgtgggac
     3601 atctgtgttc aagggaagga ctccaggagt ctgctgggaa tgaggcctag taggaaatga
     3661 ggtggccctt gagggtacag aacaggttca ttcttcgcca aattcccagc accttgcagg
     3721 cacttacagc tgagtgagat aatgcctggg ttatgaaatc aaaaagttgg aaagcaggtc
     3781 agaggtcatc tggtacagcc cttccttccc tttttttttt ttttttttgt gagacaaggt
     3841 ctctctctgt tgcccaggct ggagtggcgc aaacacagct cactgcagcc tcaacctact
     3901 gggctcaagc aatcctccag cctcagcctc ccaaagtgct gggattacaa gcatgagcca
     3961 ccccactcag ccctttcctt cctttttaat tgatgcataa taattgtaag tattcatcat
     4021 ggtccaacca accctttctt gacccacctt cctagagaga gggtcctctt gcttcagcgg
     4081 tcagggcccc agacccatgg tctggctcca ggtaccacct gcctcatgca ggagttggcg
     4141 tgcccaggaa gctctgcctc tgggcacagt gacctcagtg gggtgagggg agctctcccc
     4201 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
     4261 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
     4321 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg ggaagcttgc
     4381 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4441 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4501 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     4561 gcccaccctc gtgaccacct tcacctacgg cgtgcagtgc ttcagccgct accccgacca
     4621 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     4681 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     4741 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     4801 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     4861 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     4921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4981 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5041 catggtcctg atggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5101 caagtaaagc ggccgcgact ctaggatccg cagatcccgg ccagataccg atgctgccgc
     5161 agcaaaagca ggagcagatg ccgccgtcgc aggcgaagat gtcgcagacg gaggaggcga
     5221 tgctgccggc ggaggaggcg aagtaagtag agggctgggc tgggctgtgg ggggtgtggg
     5281 gtgcgggact gggcagtctg ggagtccctc tcaccacttt tcttaccttt ctaggatgct
     5341 gccgtcgccg ccgctcatac accataaggt gtaaaaaata ctagatgcac agaatagcaa
     5401 gtccatcaaa actcctgcgt gagaatttta ccagacttca agagcatctc gccacatctt
     5461 gaaaaatgcc accgtccgat gaaaaacagg agcctgctaa ggaacaatgc cacctgtcaa
     5521 taaatgttga aaactcatcc cattcctgcc tcttggtcct tgggcttggg gaggggtgcg
     5581 cggatgtggt tagggaacat gactggtcaa atgggaaggg cttcaaaaga attcccaata
     5641 ttgactacca agccacctgt acagatctgc ctgcaggcat gcaagcttgg cactggccgt
     5701 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5761 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5821 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     5881 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5941 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6001 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6061 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6121 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6181 g
//
