LOCUS       sequence-133484-        6925 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 6925)
  TITLE     Second Batch
REFERENCE   2  (bases 1 to 6925)
  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"
COMMENT     Sequence Label: pcDNA3-GFP-MMP7
FEATURES             Location/Qualifiers
     source          1..6925
                     /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(2477..2495)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2478..2495)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2515..2532)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2521..2745
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2791..3219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2878..2897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3088..3109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3228..3248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3233..3563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3414..3549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3476..3495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3630..4424
                     /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(3684..3703)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4294..4313
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4598..4719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4635..4654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4689..4708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4768..4784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4768..4784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4781..4803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4792..4808
                     /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(4816..4822,4823..4840,4841..4846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4861..4882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4999..5016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5170..5758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5250..5269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5929..6720,6721..6789))
                     /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     6552..6571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6790..6894)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 attccgactc accgtgctgt gtgctgtgtg cctgctgcct
     1681 ggcagcctgg ccctgccgct gcctcaggag gcgggaggca tgagtgagct acagtgggaa
     1741 caggctcagg actatctcaa gagattttat ctctatgact cagaaacaaa aaatgccaac
     1801 agtttagaag ccaaactcaa ggagatgcaa aaattctttg gcctacctat aactggaatg
     1861 ttaaactccc acgtcataga aataatgcag aagcccagat gtggagtgcc agatgttgca
     1921 gaatactcac tatttccaaa tagcccaaaa tggacttcca aagtggtcac ctacaggatc
     1981 gtatcatata ctcgagactt accgcatatt acagtggatc gattagtgtc aaaggcttta
     2041 aacatgtggg gcaaagagat ccccctgcat ttcaggaaag ttgtatgggg aactgctgac
     2101 atcatgattg gctttgcgcg aggagctcat ggggactcct acccatttga tgggccagga
     2161 aacacgctgg ctcatgcctt tgcgcctggg acaggtctcg gaggagatgc tcacttcgat
     2221 gaggatgaac gctggacgga tggtagcagt ctagggatta acttcctgta tgctgcaact
     2281 catgaacttg gccattcttt gggtatggga cattcctctg atcctaatgc agtgatgtat
     2341 ccaacctatg gaaatggaga tccccaaaat tttaaacttt cccaggatga tattaagggc
     2401 attcagaaac tatatggaaa gagaagtaat tcaagaaaga aatagaaact tcaggcagaa
     2461 ctctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2521 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2581 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2641 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2701 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2761 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2821 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2881 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2941 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3001 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3061 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3121 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3181 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3241 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3481 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3541 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3601 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3661 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3721 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3781 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3841 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3901 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3961 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4021 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4081 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4141 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4201 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4261 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4321 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4381 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4441 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4501 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4561 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4621 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4681 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4741 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4801 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4861 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4921 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4981 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5041 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5101 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5341 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5581 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6901 tccccgaaaa gtgccacctg acgtc
//
