LOCUS       sequence-483719-        8396 bp    DNA     circular SYN 18-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8396)
  TITLE     Second Batch
REFERENCE   2  (bases 1 to 8396)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pcDNA-GFP-STAT1
FEATURES             Location/Qualifiers
     source          1..8396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(922..924,925..927,928..1638)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLCYGVQCFSRYPGHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNFKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(967..988)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1575..1596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3949..3967)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3950..3967)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3987..4004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3993..4217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4263..4691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4350..4369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4560..4581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4700..4720)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4705..5034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4885..5020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4947..4966
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5101..5895
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5155..5174)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5765..5784
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6069..6190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6106..6125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6160..6179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6239..6255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6239..6255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6252..6274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6263..6279
                     /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)."
     promoter        complement(join(6287..6293,6294..6311,6312..6317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6332..6353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6470..6487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6641..7229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6721..6740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7400..8191,8192..8260))
                     /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     8023..8042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8261..8365)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgatcca
      901 gcctccgcgg gcgcgcaaga aatggctagc aagggcgagg agctgttcac cggggtggtg
      961 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1021 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1081 ctgcccgtgc cctggcccac actagtgacc accctgtgct acggcgtgca gtgcttcagc
     1141 cgctaccccg gccacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1201 gtccaggagc gcaccatctt cttcaaggac gacggcaact tcaagacccg cgccgaggtg
     1261 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1321 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1381 atggccgaca agcagaagaa cggcatcaag gttaacttca agacccgcca caacatcgag
     1441 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1501 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1561 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccggcat cactcacggc
     1621 atggacgagc tgtacaagga attctgcaga tatccatcac actggcggcc gcctcagtgg
     1681 tacgaacttc agcagcttga ctcaaaattc ctggagcagg ttcaccagct ttatgatgac
     1741 agttttccca tggaaatcag acagtacctg gcacagtggt tagaaaagca agactgggag
     1801 cacgctgcca atgatgtttc atttgccacc atccgttttc atgacctcct gtcacagctg
     1861 gatgatcaat atagtcgctt ttctttggag aataacttct tgctacagca taacataagg
     1921 aaaagcaagc gtaatcttca ggataatttt caggaagacc caatccagat gtctatgatc
     1981 atttacagct gtctgaagga agaaaggaaa attctggaaa acgcccagag atttaatcag
     2041 gctcagtcgg ggaatattca gagcacagtg atgttagaca aacagaaaga gcttgacagt
     2101 aaagtcagaa atgtgaagga caaggttatg tgtatagagc atgaaatcaa gagcctggaa
     2161 gatttacaag atgaatatga cttcaaatgc aaaaccttgc agaacagaga acacgagacc
     2221 aatggtgtga caaagagtga tcagaaacaa gaacagctgt tactcaagaa gatgtattta
     2281 atgcttgaca ataagagaaa ggaagtagtt cacaaaataa tagagttgct gaatgtcact
     2341 gaacttaccc agaatgccct gattaatgat gaactagtgg agtggaagcg gagacagcag
     2401 agcgcctgta ttggggggcc gcccaatgct tgcttggatc agctgcagaa ctggttcact
     2461 atagttgcgg agagtctgca gcaagttcgg cagcagctta aaaagttgga ggaattggaa
     2521 cagaaataca cctacgaaca tgaccctatc acaaaaaaca aacaagtgtt atgggaccgc
     2581 accttcagtc ttttccagca gctcattcag agctcgtttg tggtggaaag acagccctgc
     2641 atgccaacgc accctcagag gccgctggtc ttaaagacag gggtccagtt cactgtgaag
     2701 ttgagactgt tggtgaaatt gcaagagctg aattataatt tgaaagtcaa agtcttattt
     2761 gataaagatg tgaatgagag aaatacagta aaaggattta ggaagttcaa cattttgggc
     2821 acgcacacaa aagtgatgaa catggaggag tccaccaatg gcagtctggc ggctgaattt
     2881 cggcacctgc aattgaaaga acagaaaaat gctggcacca gaacgaatga gggtcctctc
     2941 atcgttactg aagagcttca ctcccttagt tttgaaaccc aattgtgcca gcctggtttg
     3001 gtaattgacc tcgagacgac ctctctgccc gttgtggtga tctccaacgt cagccagctc
     3061 ccgagcggtt gggcctccat cctttggtac aacatgctgg tggcggaacc caggaatctg
     3121 tccttcttcc tgactccacc atgtgcacga tgggcccagc tttcagaagt gctgagttgg
     3181 cagttttctt ctgtcaccaa aagaggtctc aatgtggacc agctgaacat gttgggagag
     3241 aagcttcttg gtcctaacgc cagccccgat ggtctcattc cgtggacgag gttttgtaag
     3301 gaaaatataa atgataaaaa ttttcccttc tggctttgga ttgaaagcat cctagaactc
     3361 attaaaaaac acctgctccc tctctggaat gatgggtgca tcatgggctt catcagcaag
     3421 gagcgagagc gtgccctgtt gaaggaccag cagccgggga ccttcctgct gcggttcagt
     3481 gagagctccc gggaaggggc catcacattc acatgggtgg agcggtccca gaacggaggc
     3541 gaacctgact tccatgcggt tgaaccctac acgaagaaag aactttctgc tgttactttc
     3601 cctgacatca ttcgcaatta caaagtcatg gctgctgaga atattcctga gaatcccctg
     3661 aagtatctgt atccaaatat tgacaaagac catgcctttg gaaagtatta ctccaggcca
     3721 aaggaagcac cagagccaat ggaacttgat ggccctaaag gaactggata tatcaagact
     3781 gagttgattt ctgtgtccga agttcaccct tctagacttc ggaccacaga caacctgctc
     3841 cccatgtctc ctgaggagtt tgacgaggtg tctcggatag tgggctctgt agaattcgac
     3901 agtatgatga acacagtata ggtcgagcat gcatctagag ggccctattc tatagtgtca
     3961 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4021 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4081 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4141 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4201 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4261 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4321 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4381 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4441 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4501 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4561 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4621 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4681 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4741 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4801 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4861 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4921 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4981 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5041 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5101 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5161 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5221 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5281 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5341 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5401 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5461 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5521 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5581 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5641 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5701 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5761 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5821 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5881 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5941 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6001 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6061 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6181 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7081 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
